Protein kinase C modulators. K.

ABSTRACT

Compounds having anti-inflammatory and other activities are disclosed. The compounds are derived from diterpenes of the phorbol-, resiniferonol- and ingenol-classes.

RELATED APPLICATIONS

This application is a continuation-in-part of: (i) application Ser. No.07/664,397, filed Mar. 4, 1991, which is a continuation-in-part of: (a)application Ser. No. 07/537,885, filed Jun. 14, 1990 (abandoned), whichis a file-wrapper continuation-in-part of application Ser. No.07/061,299, filed Jun. 10, 1987 (abandoned), which is acontinuation-in-part of application Ser. No. 06/872,812, filed Jun. 11,1986 (abandoned); and (b) application Ser. No. 07/559,296, filed Jul.30, 1990 (abandoned) and application Ser. No. 07/559,701, filed Jul. 30,1990 (now U.S. Pat. No. 5,145,842, issued Sep. 2, 1992), both of whichare a continuation-in-part of application Ser. No. 07/322,881, filedMar. 13, 1989 (abandoned), which is a division of application Ser. No.07/061,299, filed Jun. 10, 1987 (abandoned), which is acontinuation-in-part of application Ser. No. 06/872,812, filed Jun. 11,1986 (abandoned); and, (ii) application Ser. No. 07/980,907, filed Nov.24, 1992, all of which are herein incorporated by reference.

BACKGROUND

Protein kinase C (also known as "calcium/phospholipid-dependent proteinkinase", "PKC" or C-kinase") is a family of very closely relatedenzymes; one or more members of the protein kinase C family are found innearly all animal tissues and animal cells that have been examined. Theidentity of protein kinase C is generally established by its ability tophosphorylate proteins when adenosine triphosphate and phospholipidcofactors are present, with greatly reduced activity when thesecofactors are absent. Protein kinase C is believed to phosphorylate onlyserine and/or threonine residues in the proteins that are substrates forprotein kinase C. Additionally, some forms of protein kinase C requirethe presence of calcium ions for maximal activity.

Protein kinase C activity is also substantially stimulated by certain1,2-sn-diacylglycerols that bind specifically and stoichiometrically toa recognition site on the enzyme. This site is called the diacylglycerolbinding site, and it is located on the amino-terminal portion of proteinkinase C, the so-called "regulatory domain". The carboxy-terminalportion of protein kinase C carries the site at which proteinphosphorylation is effected, and this portion thus called the "kinasedomain".

Thus, the rate at which various protein kinase C family members carryout their enzymatic phosphorylation of certain substrates can bemarkedly enhanced by the presence of the cofactors such asphospholipids, diacylglycerols and, for some protein kinase C familymembers, calcium ions. This stimulation of protein kinase C activity isreferred to as protein kinase C "activation", and the activation ofprotein kinase C by the binding of diacylglycerols to the regulatorydomain of protein kinase C is of particular importance in the normal andpathological functions of protein kinase C.

In contrast to the activation of protein kinase C, some chemicalcompounds have been shown, when added to protein kinase C enzyme assays,to reduce the rate at which protein kinase C phosphorylates itssubstrates; such compounds are referred to as protein kinase C"inhibitors" or, in some cases, "antagonists". In some circumstances,protein kinase C inhibitors are capable of inhibiting various cellularor tissue phenomena which are thought to be mediated by protein kinaseC.

Activation of protein kinase C by diacylglycerols has been shown to bean important physiological event that mediates the actions of a widevariety of hormones, neurotransmitters, and other biological controlfactors such as histamine, vasopressin, alpha-adrenergic agonists,dopamine agonists, muscarinic cholinergic agonists, platelet activatingfactor, etc. [see Y. Nishizuka, Nature 308, 693-698 (1984) and Science225, 1365-1370 (1984) for reviews].

The biological role of protein kinase C is also of great interestbecause of the discovery that certain very powerful tumor promotingchemicals activate this enzyme by binding specifically and with veryhigh affinity to the diacylglycerol binding site on the enzyme. Inaddition to diacylglycerols, there are at present five other knownclasses of compounds that bind to this site: diterpenes such as thephorbol esters; indole alkaloids (indolactams) such as the teleocidins,lyngbyatoxin, and indolactam V; polyacetates such as the aplysiatoxinsand oscillatoxins; certain derivatives of diaminobenzyl alcohol; andmacrocyclic lactones of the bryostatin class. The phorbol esters havelong been known as powerful tumor promoters, the teleocidins andaplysiatoxins are now known to have this activity, and it appears likelythat additional classes of compounds will be found to have the toxic andtumor promoting activities associated with the capability to bind to thediacylglycerol site of protein kinase C and thus activate the enzyme.Other toxicities of these agents when administered to animals includelung injury and profound changes in blood elements, such as leukopeniaand neuropenia.

In addition to potent tumor promoting activity, these six classes ofcompounds, collectively referred to herein as "phorboids", display avast range of biological activities, as would be expected from thewidespread distribution of their target enzyme. Some of theseactivities, like tumor promotion, indicate the involvement of proteinkinase C in important normal or pathological processes in animals. Thus,the phorboids are potent skin inflammatory agents, cause smooth musclecontraction in several tissues, alter immune system function and can beused to cause a variety of other normal or pathological responses.Related disease states such as the development of cancer, the onsetand/or maintenance of inflammatory disease, the role of vasoconstrictionin hypertension, the role of bronchoconstriction in asthma, the lifecycles of many pathogenic human viruses, and the role of cholinergic,adrenergic, and dopaminergic synapses in diseases of thecentral/peripheral nervous systems, may be mediated in vivo by thestimulation of protein kinase C or other diacylglycerol bindingsite-bearing entities by diacylglycerols, the latter being generated inthe cell by pathological agents or conditions.

In analyzing the activity of a pharmaceutical or other bioactivecompound, it is useful to consider two properties: the efficacy, definedas the capability to elicit a full or partial biological result, such ascomplete displacement of a ligand from its receptor site or the completeinhibition of inflammation or edema caused by a standard stimulus; andthe potency, defined as that amount or concentration of drug that causes50% of the full response (often abbreviated as the ED₅₀). It isfrequently the case within a given class of pharmaceutical agents thatindividual members of the class all have equal efficacy, i.e. they eachcan generate a full biological effect, but they show differingpotencies. Thus, the structural modifications within such a class affectonly the amount necessary to achieve a given result, and the modifiedcompounds otherwise have generally the same central biologicalcharacteristic. There may also be differences between members of such aclass as regards properties other than the central biologicalcharacteristic; for example, members of the class might differ in sideeffects or toxicity.

Well-known pharmaceuticals that have been in extensive use for years ordecades show a wide range of optimal therapueutic potencies. Aspirin,for example, is often taken in multi-gram amounts per day for treatmentof inflammation or arthritis, and detailed analyses of its mechanism ofaction in vitro show that a concentration in the millimolar range isrequired. In contrast, steroid-based topical anti-inflammatory compoundssuch as fluocinolone acetonide are many thousand-fold more potent, and,beyond this, some oral contraceptive agents are prescribed in dailydoses in the microgram range. Thus, although high potency is generallyadvantageous for a pharmaceutical, it is not an absolute requirement.

A thousand or more analogs of the highly skin-inflammatory andtumor-promoting phorboids have been reported in the literature,including numerous examples on which minor chemical modifications havebeen made [see Evans and Soper, Lloydia 41, 193-233 (1978) andreferences cited therein]. The structures of these phorboids can becompared, and their activities for inflammation and tumor promotion canbe analyzed from the perspective of efficacy and potency. The structuresof the different classes of phorboids vary quite markedly from one tothe other class (see Summary of the Invention for structuralcomparisons), yet widespread testing of their biological activities hasshown that these classes have generally very similar biologicalproperties. In particular, the thousands of known phorboids of thehighly potent diterpene, indolactam, and polyacetate classes appear tohave, with very minor exceptions, virtually identical efficacies as skinirritants and tumor promoters [T. Sugimura, Gann 73, 499-507 (1982)].The exceptions involve a few compounds that have a short duration ofirritant activity and/or manifest diminished tumor promoting activity,perhaps due to toxicity or secondary parameters such as differingmetabolic destruction rates.

In contrast to the essentially equal efficacies among the vast majorityof phorboids, their relative potencies cover a wide range, as measuredin inflammation and promotion tests and as measured in numerous other invivo and in vitro systems. Example compounds can be found in thediterpene, indolactam, and polyacetate classes that have nearly equal,very high potencies. At the same time there are compounds in each ofthese classes which embody significant structural changes that do notdiminish efficacy but do result in potency decreases of 10-fold to100,000-fold or more [see, for example, Driedger and Blumberg, CancerRes. 37, 3257-3265 (1977), Cancer Res. 39, 714-719 (1979)]. Thus, allthese compounds appear to be capable of achieving generally the samebiological results, and merely differ in the amount which must be usedto obtain a given result.

In vitro measurements of biochemical properties provide an even moresensitive method for comparing the properties of the various phorboids.For example, using a radioactively labeled phorboid such as [³ H]phorbol12,13-dibutyrate or [³ H]lyngbyatoxin, one can measure the potency of atest compound as a competitive ligand for the diacylglycerol bindingsite, which is also referred to herein as the "phorboid binding site" onprotein kinase C or on other biological molecules which have phorboidbinding sites (see below). Alternatively, one can measure the ability ofa given phorboid to stimulate the protein kinase C-mediatedincorporation of radioactive phosphate from [³² P]adenosine triphosphateinto a standard acceptor substrate such as histone H1. Tests of thisnature reveal a difference in potency between given phorboid agonists ofas much as 10,000,000-fold or more [Dunn and Blumberg, Cancer Res. 43,4632-4637 (1983), Table 1].

These basic data regarding the phorboid agonists are an importantconsideration because they underscore the concept that the structuraldifferences among these previously known phorboids, especially thediterpenes, indolactams, polyacetates, and bryostatins, generally do notaffect their efficacies as toxic agonists, and indeed a wide variety ofstructural changes are tolerated in this regard. Such changes generallyalter potency only and do not provide agents with therapeutic utility,since they retain their toxicity.

Some minor changes in phorboid structure are known to result in inactivecompounds, such as a stereochemical change from 4-beta to 4-alpha in thephorbol series, and indeed some of the diterpene skeleton structurescarry hydroxy groups that must be esterified in order for inflammatoryactivity to be observed. However, these inactive compounds are quite fewin number among the known phorboids, and no therapeutic utility has beendemonstrated for them.

The phorbol esters, indolactams, polyacetates, diaminobenzyl alcohols,and bryostatins are generally found in plants, molds, and algae, or aresynthetic in origin. Although they are found in many parts of the world,normal human contact with them is thought to be low. In contrast, thediacylglycerols are part of the functioning of virtually every type ofanimal cell except erythrocytes of some species, and thus theundesirable activation of protein kinase C by the diacylglycerols mayhave a very widespread role in human diseases.

Thus, compounds capable of blocking the activation of, or inhibiting,protein kinase C by acting as specific pharmacological antagonists ofthe diacylglycerols at the diacylglycerol binding site on protein kinaseC, would be valuable agents in the prevention and treatment of a widevariety of diseases in animals and humans. For example, the need for,and potential utility of, protein kinase C inhibitors/antagonists asagents for the treatment of cancer has received much attention [D.Corda, et al., Trends in Pharmacological Sciences 11, 471-473 (1990); G.Powis, Trends in Pharmacological Sciences 12, 188-194 (1991); S. Gandyand P. Greengard, Trends in Pharmacological Sciences 13, 108-113 (1992);B. Henderson and S. Blake, Trends in Pharmacological Sciences 13,145-152 (1992)].

Protein kinase C comprises a family of eight or more closely relatedprotein molecules [Parker, P. J. et al. Mol. Cell. Endocrin. 65. 1-11(1989)]. Because of their high degree of relatedness they are referredto as "isozymes", "isotypes" or "isoforms". Occasionally the term"subtypes" is used, but this term is usually reserved to designate, as asubdivision, two or more variants of a single isotype.

The known isotypes of protein kinase C are α, β₁, β₂ and γ (the"A-group"), δ, ε, ε'[Ono, Y. et al., J. Biol. Chem. 263, 6927-6932(1988), ζ, and the recently described "protein kinase C-L" [Bacher, N.et al., Mol. Cell. Biol. 11,126-133 (1991)], also known as proteinkinase Cη [sada, S. et al., J. Biol. Chem. 265, 22434-22440 (1990)] (the"B-Group"). Members of the A-group require calcium ions for maximalactivation, whereas the B-group members are thought to be largelycalcium-independent for activation. The genes for each of the isotypesabove have been cloned from one or more animal and yeast species and theclones have been sequenced; thus the relatedness of the genes and theirproduct polypeptides is thus well established. A possible new,as-yet-unnamed member has also recently been described [Bignon, E. etal., Biochem. Biophys. Res. Comm. 171, 1071-1078 (1990)], but no DNA orprotein sequence information is available to confirm the novelty of thisprotein kinase C-like activity.

It is possible that the different protein kinase C isozymes havedifferent biological roles, and published evidence supports this idea[Homan, E., Jensen, D. and Sando, J., J. Biol. Chem. 266, 5676-5681(1991); Gusovsky, F. and Gutkind, S., Mol. Pharm. 39. 124-129 (1991);Borner, C., "The Role of protein kinase C in Growth Control", SixthInternational Symposium on Cellular Endocrinology, W. Alton Jones CellScience Center, Lake Placid, N.Y., Aug. 12-15, 1990; Naor, Z. et al.,Proc. Natl. Acad. Sci. USA 86, 4501-4504 (1989); Godson, C., Weiss, B.and Insel, P., J. Biol. Chem. 265, 8369-8372 (1990); Melloni, E. et al.,Proc. Natl. Acad. Sci. USA. 87, 4417-4420 (1990); Koretzky, G. et al.,J. Immunology 143, 1692-1695 (1989)]. For example, the stimulation ofone protein kinase C isotype or a limited subset of protein kinase Cisotypes might lead to undesirable results such as the development ofinflammation [Ohuchi, K. et al., Biochim. Biophys. Acta 925, 156-163(1987)], the promotion of tumor formation [Slaga, T., Envir. HealthPerspec. 50, 3-14(1983)] or an increased rate of viral replication incells (i.e., de novo infection of cells and/or expression, assembly andrelease of new viral particles) [Harada, S. et al., Virology 154,249-258 (1986)].

On the other hand, other protein kinase C isozymes might be responsiblefor the many beneficial effects observed when protein kinase C isstimulated by known protein kinase C activators in a variety ofbiological settings; such beneficial effects include the cessation ofdivision of leukemic cells [Rovera, G., O'Brien, T. and Diamond, L.,Science 204, 868-870 (1979)], multiplication of colonies of lymphocytes[Rosenstreich, D. and Mizel, S., J. Immunol. 123, 1749-1754 (1979)] andleucocytes [Skinnider, L. and McAskill, J., Exp. Hematol. 8, 477-483(1980)] or the secretion of useful bioregulatory factors such asinterferon-γ [Braude, I., U.S. Pat. No. 4,376,822] and interleukin-2[Gillis, S., U.S. Pat. nO. 4,401,756].

Recent publications indicate that diacylglycerol binding sites exist onnewly-described proteins which lack the kinase domain, and thus lack thekinase activity, of protein kinase C. One such protein is n-chimaerin,found in human brain [Ahmed et al., Biochem. J. 272, 767-773 (1990)] andthe other is the unc-13 gene product of the nematode Caenorhabditiselegans, [Maruyama, I. and Brenner, S., Proc. Natl. Acad. Sci. USA 815,5729-5733 (1991)]. The presence of the diacylglycerol binding sites onthese two proteins was demonstrated by standard binding experiments with[³ H]phorbol 12,13-dibutyrate. These new proteins may have otherenzymatic or biological activities which can be modulated by compoundswhich bind to their diacylglycerol binding sites. Thus, such compoundsmay have utility on non-protein kinase C biological targets.

Given that there are now numerous distinct biological entities bearingdiacylglycerol binding sites, it would be highly desirable to obtainchemical compounds which could specifically and selectively target oneor another type of diacylglycerol binding site, thus permitting one toselectively activate or inhibit one such site without affecting theothers. Such compounds would be valuable experimental tools for studyingthe role of individual types of proteins bearing diacylglycerol bindingsites as well as providing novel means for treating diseases in whichprotein kinase C or other diacylglycerol binding site-bearing proteinsare involved.

There are several published reports describing chemical compoundscapable of selectively distinguishing severaldiacylglycerol/phorboid-type binding sites in mouse skin [Dunn andBlumberg, op. cit.] and in purified preparations of protein kinase Cisotypes [Ryves, W. J. et al., FEBS Letters 288, 5-9 (1991)]. However,in these studies, even the compounds showing the clearest differences inaffinity for these distinct classes, namely phorbol 12,13-dibutyrate,12-deoxyphorbol 13-isobutyrate, 12-deoxyphorbol 13-phenylacetate andthymeleatoxin, are only selective by a factor of 10-1000 in dissociationconstant among the different binding sites. Furthermore, these compoundshave potent skin inflammatory activity and are not desirable in human oranimal medicine because of this toxicity.

Thus, to briefly recapitulate, two kinds of new compounds relating todiacylglycerol binding sites would be highly desirable. The first typewould be capable of selectively activating one or a few useful, but notother, deleterious, diacylglycerol target sites. The second type wouldbe capable of inhibiting, or antagonizing the stimulation of, one ormore deleterious diacylglycerol binding site-bearing entities withoutblocking the useful ones. These kinds of compounds would be valuableagents for the study of diacylglycerol binding site-bearing entities andfor the prevention or treatment of a wide range of human and animaldiseases thought to involve protein kinase C or other entities under thecontrol of diacylglycerol binding sites.

Earlier efforts to use the previously known phorboids themselves or tomodify the structures of these known phorboids, have generally not beensuccessful in producing useful compounds with toxicity low enough foruse in humans.

It has been known for some time that several of the toxic, inflammatoryand tumor-promoting compounds such as phorbol 12-tigliate 13-decanoate,mezerein, lyngbyatoxin and aplysiatoxin have anti-leukemic activity inmouse model tests [T. Sugimura, op cit.; S. M. Kupchan and R. L. Baxter,Science 187, 652-653 (1975); S. M. Kupchan, et al., Science 191, 571-572(1976); M. C. Territo and H. P. Koeffler, Br. J. Haematol. 47, 479-483(1981)]. However, these compounds are all extremely toxic and are cancersuspect agents, thus eliminating them from consideration as humantherapeutic agents.

Ganong et al. [Proc. Nat. Acad. Sci. USA 83, 1184-1188 (1986)] tested aseries of diacylglycerols and found no antagonistic activity in thatseries against the standard agonist, 1,2-dioctanoylglycerol. It is ofparticular note that several compounds tested in this work were modifiedin the hydroxymethyl portion of the diacylglycerol molecule, and thesemodifications produced only a loss of activity or a weakened activitythat was not distinguishable from the agonist activity of1,2-dioctanoylglycerol itself, a compound which is toxic to mouse skin[R. Smart, et al., Carcinogenesis 7, 1865-1870 (1986); A. Verma, CancerRes. 48, 2168-2173 (1988)]. These hydroxymethyl-modified compounds werenot antagonists in these tests and no utility was found. Similarly,Thielmann and Hecker [Forsch. Krebsforsch. Vol. VII, pp. 171-179 (1969),New York: Schattauer] found only a complete loss of biological activityin their study when the hydroxy group of the hydroxymethyl on phorbol12,13-didecanoate was replaced with hydrogen or chlorine. Schmidt andHecker [H. Lettre and G. Wagner (eds.), Aktuelle Probleme aus dem Gebietder Cancerologie, Vol. III, 3rd Heidelberg Symposium, pp. 98-108.Berlin: Springer Verlag, 1971.] also found that oxidation of thehydroxymethyl of phorbol 12,13- didecanoate to a carboxylic acid causedcomplete loss of activity in the assays used.

The hydroxymethyl group of the known phorboids (discussed in more detailin Summary of the Invention) has been thought to be required forbiological activity, as detailed by Hecker (Hecker, E., Carcinogenesis,Vol. 2, eds. Slaga, Sivak and Boutwell, Raven Press, New York, 1978, pp.11-48 and references cited therein). Indeed, it is stated therein thatthe replacement of the 20-hydroxyl in a phorbol ester "results incomplete loss of biological activity". In another study, replacement ofthe hydroxy group of the hydroxymethyl (located at carbon 14) bychlorine or hydrogen in indolactam V gave rise to compounds with agonistactivity weaker than but otherwise not distinguished from the agonistactivity of the very toxic teleocidin class of tumor promoters [Irie etal., Int. J. Cancer 36, 485-488 (1985)]. Thus no utility beyond that ofthe toxic, hydroxymethyl-bearing parent indolactam-type compounds wasfound.

Schmidt and Hecker (Carcinogenesis, Vol. 7, ed. by E. Hecker et al.,Raven Press, New York, 1982, pp. 57-63) studied the abilities of aseries of diterpene phorboids to inhibit tumor promotion by the standardphorboid agonist tumor promoter phorbol 12-myristate 13-acetate (PMA).They found that, at low doses, some short-chain ester derivatives ofphorbol were able to block the tumor promotion by PMA. However, all ofthe compounds that were active as antagonists at low doses are also veryefficacious skin irritants themselves at slightly higher doses and mostof them are also known to have tumor promoting activity. Thus, theseshort-chain esters still have toxic inflammatory and tumor promotingactivity at doses only slightly different from those which would beneeded to exhibit a therapeutic effect in mice.

SUMMARY OF THE INVENTION

This invention pertains to novel phorboid derivatives which variouslyblock the toxic effects of the hydroxymethyl-containing phorboids, lackthe toxic properties of previously available phorboids and show activityfor applications as therapeutics. The phorboid derivatives of thepresent invention embody very diverse structures and have utility asanti-inflammatory agents, as cancer cell and leukemic cell inhibitoryagents, anti-asthmatic and anti-hypertensive agents, as modulators ofhuman immune cell function, as anti-viral agents, as stimulators of theproduction of lymphokines such as interferon and the interleukins, ascentral nervous system pharmaceuticals for several pathologicalconditions, and as xenobiotics for achieving the control of parasites.

The structural features associated with the non-toxicity anddiacylglycerol binding site modulating properties of these compoundsrelate primarily to the hydroxymethyl or 1-hydroxyethyl group found ineach of the toxic parent compounds. Specific modifications of the latterchemical groupings yields non-skin inflammatory compounds that showanti-inflammatory activity in several test systems, whereas any of avery wide variety of changes in other parts of the parent phorboidstructures, including but not limited to diterpenes, indole alkaloids,polyacetates, diaminobenzyl alcohol derivatives, aplysiatoxins, andbryostatinoids, have very markedly less effect on the overall biologicalproperties of the derivatives, other than changes in potency. Thisinvention also provides new compounds that discriminate between phorboidreceptor-type targets, with direct or imputed relative bindingactivities differing by 10,000-fold or more in some cases.

A comparison of the structures of the previously known phorboids revealsthe features that are critical for this invention, as shown below, inwhich the hydroxymethyl and 1-hydroxyethyl groups are enclosed in dashedlines. ##STR1## It can be seen that the phorboids depicted have diversestructural elements of both hydrophilic and hydrophobic nature, with oneprominent exception, namely that each contains a hydroxymethyl or1-hydroxyethyl group (indicated by the dashed-line boxes in eachstructure). In each case the phorboid depicted is among the most potentof its particular structural class, and among the classes thediterpenes, indolactams, polyacetates and bryostatins have members ofespecially high potency.

The hydroxymethyl and 1-hydroxyethyl feature common to all the classesof phorboids is the primary focus of this invention, in which thevariations which can be accommodated in the organic functional groupsreplacing the hydroxymethyl/1-hydroxyethyl groups are found to includenew heteroatoms and to be larger and more complex than previouslyrecognized. Furthermore, the parent structures ofhydroxymethyl/1-hydroxyethyl-modified phorboids have now also been foundto tolerate larger and more complex structural features and more kindsof heteroatoms than previously recognized.

Generally the phorboid derivatives of this invention can be representedby the formula:

    P--G

wherein P represents a radical, formally derived from a parent compound,which compound:

a. binds reversibly or irreversibly to a diacylglycerol-type receptor;and/or

b. activates any form of the enzyme protein kinase C; and

c. contains an hydroxymethyl or 1-hydroxyethyl group bonded to a carbonatom; and

wherein G is any group of 55 or fewer atoms selected from carbon,hydrogen, oxygen, nitrogen, halogen, sulfur, phosphorus, silicon,arsenic, boron and selenium either i) singly or doubly bonded to thecarbon atom of the parent compound in place of the hydroxymethyl or1-hydroxyethyl group; or ii) singly or doubly bonded to a carbon atomimmediately adjacent to the carbon atom to which the hydroxymethyl or1-hydroxyethyl group is bound in the parent compound; and wherein thehydroxymethyl or 1-hydroxyethyl group of the parent compound is absentor has been replaced by G.

More specifically, the compounds of this invention are represented bythe formula:

    P.sub.o --S.sub.o --E.sub.o

which depicts a phorboid "parent" compound radical P_(o) modified by asubstituent S_(o) E_(o).

In general, a parent compound is a compound which:

a. binds reversibly or irreversibly to a diacylglycerol-type receptor;and/or

b. activates any form of the enzyme protein kinase C; and

c. contains an hydroxymethyl or 1-hydroxyethyl group bonded to a carbonatom.

For example, a parent compound can be a diterpenoid activator of proteinkinase C; an aromatic heterocyclic activator of protein kinase of theindole, indene, benzofuran, or benzothiophene class, further definedhere by the presence of a substituted or unsubstituted six-atom chainconnecting positions 3 and 4 of the indole, indene, benzofuran orbenzothiophene to form an additional 9-membered ring and by the optionalpresence in this class of one or two nitrogen atoms at any of positions5, 6 and 7 of the benzenoid ring portion of the indole, indene,benzofuran or benzothiophene skeletons; a polyacetate-derived activatorof protein kinase C; an activator of protein kinase C of thediacylglycerol or diacyloxybutanol class; an activator of protein kinaseC of the diaminobenzyl alcohol class; or a protein kinase C activator ofthe bryostatin class. The word "chain", when used in describing thecompounds of this invention, means a sequence of atoms covalently bound,one to the next. These sequences of covalently bound atoms may besubstituted or unsubstituted as further specified in the particularsdescribed herein.

S_(o) --E_(o) represents a modifying substituent which is either:

i) singly or doubly bonded to the carbon atom of the parent compoundP_(o) in place of the hydroxymethyl or 1-hydroxyethyl group; or

ii) singly or doubly bonded to a carbon immediately adjacent to thecarbon atom to which the hydroxymethyl or 1-hydroxyethyl is bonded.

In the S_(o) --E_(o) substituent, S_(o) can be a substituted orunsubstituted, saturated, unsaturated and/or aromatic, straight orbranched, acyclic, ring-containing and/or ring-carrying chain of atomswhich separates P_(o) and E_(o) by a linear count of at least two butnot more than 12 atoms and contains and/or carries not more than 9heteroatoms selected from oxygen, nitrogen, silicon, sulfur, phosphorus,arsenic, boron and selenium, and not more than 16 halogen atoms;provided that the total number of atoms does not exceed 35; and in somecases S_(o) may be a single or double bond; and E_(o) can be hydrogen,halogen or a saturated or singly or multiply unsaturated groupcontaining up to 15 carbon atoms and optionally containing 1 to 12halogen atoms and/or 1 to 6 heteroatoms selected from oxygen, nitrogen,silicon, sulfur, phosphorus, arsenic, boron and selenium. S_(o) E_(o)taken together may also be a hydrogen, halogen, thionic sulfur atom orketonic oxygen atom or a hydroxy, amino, or thiol group singly or doublybonded to the carbon atom of the parent compound P_(o) in place of thehydroxymethyl or 1-hydroxyethyl group.

DETAILED DESCRIPTION OF THE INVENTION

All six classes of the known phorboids have one structural element incommon. Each prototypical member of these classes has either ahydroxymethyl group or a 1-hydroxyethyl group. The design of thephorboid derivatives of this invention is based on the finding that thehydroxymethyl and 1-hydroxyethyl groups, which previously were thoughtto be required for biological activity of phorboids containing thesegroups, can be replaced by other substituents of very diverse nature,even though these new substituents are very substantially larger and ofmore diverse structure and heteroatom composition than the originalhydroxymethyl/1-hydroxyethyl group.

For example, the hydroxymethyl group of a typical diterpene phorboidsuch as phorbol-12-myristate 13-acetate may even be replaced by atetracyclic structure containing some 36 carbon atoms, to form thedimeric molecule ##STR2## which still exhibits diacylglycerol sitebinding activity and anti-viral properties.

The compounds resulting from the hydroxymethyl changes described herevariously block the toxic effects of the hydroxymethyl-containingphorboids, lack the skin-inflammatory properties associated with thepreviously available phorboids and show useful activity as therapeuticagents. These new compounds thus have utility as, variously,anti-inflammatory agents, anti-viral agents and anti-leukemic agents,for example.

Although the replacement of the hydroxymethyl group or 1-hydroxyethylgroup is very specific and leads to an extreme and profound change inbiological properties, a wide range, much wider than previouslyrecognized, of structural alterations in thenon-hydroxymethyl/1-hydroxyethyl portions of the novel compounds can betolerated without material loss of their basic, favorable biologicalproperties.

The phorboid derivatives of this invention are generally represented bythe formula:

    P--G

The formula depicts a radical, P, derived from a parent compound, whichcompound:

a. binds reversibly or irreversibly to a diacylglycerol-type receptor;and/or

b. activates any form of the enzyme protein kinase C; and

c. contains an hydroxymethyl or 1-hydroxyethyl group bonded to a carbonatom; and

wherein G is any group of 55 or fewer atoms selected from carbon,hydrogen, oxygen, nitrogen, halogen, sulfur, phosphorus, silicon,arsenic, boron and selenium either i) singly or doubly bonded to thecarbon atom of the parent compound in place of the hydroxymethyl or1-hydroxyethyl group; or ii) singly or doubly bonded to a carbon atomimmediately adjacent to the carbon atom to which the hydroxymethyl or1-hydroxyethyl group is bound in the parent compound; and wherein thehydroxymethyl or 1-hydroxyethyl group of the parent compound is absentor has been replaced by G.

More specifically, the phorboid derivatives of this invention arerepresented by the formula:

    P.sub.o --S.sub.o --E.sub.o

The formula depicts a radical P_(o), formally derived from a parenthydroxymethyl-containing phorboid compound, bonded to an S_(o) --E_(o)substituent.

P_(o) represents a radical formally derived from a compound whichcontains an hydroxymethyl (or the equivalent 1-hydroxyethyl) group andwhich binds reversibly or irreversibly to a diacylglycerol-type receptorand/or activates any form of the enzyme protein kinase C. P_(o) may beformally derived from phorboids from any of the six classes listedbelow:

i) a diterpenoid activator of protein kinase C;

ii) an aromatic heterocyclic activator of protein kinase of the indole,indene, benzofuran, or benzothiophene class, further defined here by themandatory presence of a substituted or unsubstituted six-atom chainconnecting positions 3 and 4 of the indole, indene, benzofuran orbenzothiophene skeleton to form an additional 9-membered ring and by theoptional presence in this class of one or two nitrogen atoms at any ofpositions 5, 6 and 7 of the benzenoid ring portion of the indole,indene, benzofuran or benzothiophene skeletons;

iii) a polyacetate-derived activator of protein kinase C;

iv) an activator of protein kinase C of the diacylglycerol ordiacyloxybutanol class;

v) an activator of protein kinase C of the diaminobenzyl alcohol class;and

vi) a protein kinase C activator of the bryostatin class.

All of these phorboids contain an hydroxymethyl or 1-hydroxyethyl groupwhich is known to be associated with their toxic biological activity,such as skin inflammatory activity measured on the mouse ear.

S_(o) --E_(o) represents a substituent which is either:

i) singly or doubly bonded to the carbon atom of the parent compound inplace of the hydroxymethyl or 1-hydroxyethyl group; or

ii) singly or doubly bonded to a carbon immediately adjacent to thecarbon atom to which the hydroxymethyl or 1-hydroxyethyl group is boundin the parent compound.

In this S_(o) --E_(o) substituent, S_(o) can be a substituted orunsubstituted, saturated, unsaturated and/or aromatic, straight orbranched, acyclic, ring-containing and/or ring-carrying chain of atomswhich separates P_(o) and E_(o) by a linear count of at least two butnot more than 12 atoms and contains and/or carries not more than 9heteroatoms selected from oxygen, nitrogen, silicon, sulfur, phosphorus,arsenic, boron and selenium, and not more than 16 halogen atoms;provided that the total number of atoms does not exceed 35; and in somecases S_(o) may be a single or double bond; and E_(o) can be hydrogen,halogen or a saturated or singly or multiply unsaturated groupcontaining up to 15 carbon atoms and optionally containing 1 to 12halogen atoms and/or 1 to 6 heteroatoms selected from oxygen, nitrogen,silicon, sulfur, phosphorus, arsenic, boron and selenium. S_(o) E_(o)taken together may also be a hydrogen, halogen, thionic sulfur atom orketonic oxygen atom or a hydroxy, amino, or thiol group singly or doublybonded to the carbon atom of the parent compound P_(o) in place of thehydroxymethyl or 1-hydroxyethyl group.

In a preferred embodiment, the phorboid derivatives of this inventionare represented as follows:

    P.sub.x --S.sub.x --E.sub.1

wherein P_(x) can be selected from six different classes of compoundsdesignated P₁ -P₆ and defined below, wherein S_(x) is selected fromseven different structural types as defined below and E₁ is as definedbelow.

P₁, P₂, P₃, P₄, P₅, and P₆ represent compounds of each of the sixclasses of known phorboids and are defined by the formulae below.

P₁ is a radical of the formula: ##STR3## in the form of an individualisomer, an isomer mixture, a racemate or optical antipode, or apharmaceutically acceptable salt thereof; wherein A¹ and A² may beindependently selected from hydrogen, halogen and a substituent havingnot more than 34 carbon atoms, not more than 24 halogen atoms and notmore than 9 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur, or A¹ and A² taken together may complete a 5- or6-membered saturated or unsaturated carbocyclic or heterocyclic ring,optionally substituted by 1-8 halogens and/or other groups, whichhalogens and groups taken together contain a total of not more than 30carbon atoms, not more than 24 halogen atoms and not more than 9heteroatoms selected from oxygen, nitrogen, silicon, phosphorus andsulfur; A³ is a three atom chain which carries S_(x) E₁ and completes a7-membered saturated or unsaturated carbocyclic ring optionallysubstituted by 1-6 halogens and/or other groups, which halogens andgroups taken together, excluding S_(x) E₁, contain not more than 12carbon atoms, not more than 8 halogen atoms, and not more than 5heteroatoms selected from oxygen, nitrogen, silicon, phosphorus andsulfur; provided that, including S_(x) E₁, the middle carbon atom of A³is not substituted by hydroxymethyl or 1-hydroxyethyl; A⁴ completes a 6-or 7-membered carbocyclic or heterocyclic ring which is connected in theβ configuration to either carbon atom 9 or 10 and carries an 11-methylgroup in the α or β configuration, wherein A⁴ is optionally substitutedfurther by 1-10 halogens and/or other groups, the group or groupsoptionally completing 1-3 additional rings through bonds amongthemselves and/or 1-5 additional rings when taken together with A¹, A²,a ring formed by A¹ and A² together, and/or a bond to carbon atom 9,which halogens and groups taken together include not more than 40 carbonatoms, not more than 24 halogen atoms, and not more than 15 heteroatomsselected from oxygen, nitrogen, silicon, phosphorus and sulfur; J¹ ishydrogen, fluoro, chloro, hydroxy, amino, mono- or di(lower-alkyl)amino,methyl, ethyl, vinyl, ethynyl, propargyl, cyano, methoxy, ethoxy,trifluoromethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,acetoxy, propanoyloxy, acetyl, propanoyl, hydroxyacetyl,2-hydroxypropanoyloxy, 3-hydroxypropanoyl, acetamido, propanamido,hydroxyacetamido, 2-hydroxypropanamido, or 3-hydroxypropanamido (each ofwhich must be situated in the beta configuration), or J¹ taken togetherwith A¹, A², A³ or a ring formed by A¹ together with A² completes a 3-to 7-membered substituted or unsubstituted carbocyclic or heterocyclicring, the substituents of which contain not more than 15 carbon atoms,not more than 10 halogens, and not more than 8 heteroatoms selected fromoxygen, nitrogen, silicon, phosphorus and sulfur; J² is selected fromhydrogen, methyl, ethyl, hydroxymethyl, hydroxyethyl, vinyl, ethynyl,allyl, propargyl, n-propyl and isopropyl; provided that, if A¹ and A²are not linked to form a ring, A⁴ must carry a cyclopropyl at positions13 and 14; the total P₁ ring skeleton may not comprise any of thefollowing six frameworks, which are derived from various homo-, nor- andhomo-nor-steroids: ##STR4## and provided that: for all P₁, when S_(x) E₁are taken together and bonded to carbon 6, they may not comprise --CH₃,--CH═O, --CH₂ O-R_(es) wherein R_(es) is an acyl group, --CH₂ O-R_(et)wherein R_(et) is a C₁ -C₆ hydrocarbon radical or is --C(CH₃)₂ O--linked via the oxygen atom to carbon 5 of P₁, or --CH═NNHR_(h) whereinR_(h) is a nitrophenyl ring optionally bearing additional substituents;P₁ S_(x) E₁ may not include 12-beta-13-alpha-diacetoxy derivatives ofcompounds having the exact 20-carbon tigliane or 19-carbon20-nor-tigliane skeleton; P₁ S_(x) E₁ may not comprise20-deoxy-20-chlorocrotophorbolone nor 20-deoxy-20-chlorophorbol12,13-diesters wherein the ester groups are both selected from saturatedor unsaturated alkanoyl or are both benzoyl; if S_(x) E₁ is ═CH₂ bondedto carbon 6, P₁ may not carry a ring formed by --OC(CH₃)₂ O-- bonded inthe beta configuration to carbons 3 and 4; and P₁ S_(x) E₁ may notcomprise 6-deshydroxymethyl-6-carboxyphorbol 12,13-didecanoate or6-deshydroxymethyl-6-carboxyphorbol 12,13-diacetate.

P₂ is a radical of the formula: ##STR5## in the form of an individualisomer, an isomer mixture, a racemate or optical antipode, or apharmaceutically acceptable salt thereof, wherein B¹ completes a6-membered aromatic ring which may be carbocyclic or may optionallycontain one or two nitrogen atoms at any of positions 5, 6 and 7 of thering, wherein positions 5, 6 and/or 7 of B¹ are optionally andindependently substituted on carbon by halogen(s) and/or by straightchain or branched chain, cyclic or acyclic, saturated, unsaturatedand/or aromatic carbon- and/or heteroatom-containing groups, whichhalogen(s) and groups taken together contain not more than 40 carbonatoms, not more than 24 halogen atoms, and not more than 9 heteroatomsselected from oxygen, nitrogen, silicon, phosphorus and sulfur, thegroups being optionally connected to one another and/or to B² to form1-3 additional rings; B² is selected from oxygen, sulfur, sulfoxide,sulfone, monofluoromethylene, difluoromethylene, and a carbon ornitrogen atom optionally substituted by groups having not more than 15carbon atoms, not more than 24 halogen atoms, and not more than 6heteroatoms selected from oxygen, nitrogen, silicon, phosphorus andsulfur, and B² may be linked to B¹ or K¹ to form an additionalcarbocyclic or heterocyclic ring; K¹ is hydrogen, halogen or a groupcontaining not more than 15 carbon atoms, not more than 18 halogenatoms, and not more than 6 heteroatoms selected from oxygen, nitrogen,silicon, phosphorus and sulfur, and K¹ may be linked to B² or B³ or toboth B² and B³ to form one or more additional carbocyclic and/orheterocyclic rings; B³ is a 2-carbon chain optionally substituted byhalogen(s) and/or one or more groups which, taken together but excludingS_(x) E₁, contain not more than 12 carbon atoms, not more than 6 halogenatoms, and not more than 6 heteroatoms selected from oxygen, nitrogen,and sulfur; provided that, including S_(x) E₁, the carbon atom of B³bonded to K² as defined below does not carry --CH₂ OH or --CHCH₃ OH; K²is selected from oxygen, sulfur --NK⁶ or --CK⁶ K⁷ wherein K⁶ ishydrogen, hydroxy, methyl, ethyl, fluoro, n-propyl, allyl, or propargyl,and K⁷ is hydrogen, methyl, ethyl, halogen, trifluoromethyl or cyano; K³and K⁴ may be the same or may differ and each may independently behydrogen, halogen, a substituent group, or may complete an additionalring connecting K³ and K⁴ or connecting either K³ or K⁴ to K⁵, such thatK³ and K⁴ taken together contain not more than 18 carbon atoms, not morethan 24 halogen atoms, and not more than 6 heteroatoms selected fromoxygen, nitrogen, silicon, phosphorus and sulfur; K⁵ is selected fromoxygen, sulfur, sulfoxide, sulfone or --NK⁸ --, --NOK⁸ -- or --CK⁸ K⁹ --wherein K⁸ is hydrogen or a group containing not more than 30 carbonatoms, not more than 24 halogen atoms, and not more than 8 heteroatomsselected from oxygen, nitrogen, silicon, phosphorus and sulfur, and K⁹is hydrogen, methyl, ethyl, n-propyl, hydroxy, halogen, allyl,propargyl, cyano, or trifluoromethyl; provided that P₂ S_(x) E₁ may notcomprise (-)-1,10,14-O-trimethylindolactam V, and provided that if B¹completes an unsubstituted or substituted carbocyclic aromatic ring, and

B² is --NH--, --N--(C₁ -C₁₂ linear or branched alkyl or alkanoyl)--,--N--COOCH₂ C₆ H₅ -- or --N--COOC(CH₃)₃ --, and B³ is --CH₂ CH--, and

K¹ is hydrogen, and

K² is --NH--, and

K⁴ is hydrogen, and

K⁵ is --NH-- or --N(C₁ -C₃ -alkyl)--,

then (i) if S_(x) E₁ is bonded to the carbon atom in B³ that is adjacentto K², then S_(x) E₁ may not be --COOMe or --COOEt; and (ii) if S_(x) isa single bond directed to the carbon atom in B³ that is adjacent to K²,and E₁ is --CH₂ --R_(e) ^(e), then R_(e) ^(e) is not any of hydrogen,chloro, bromo, C₁ -C₁₂ saturated or unsaturated linear or branchedalkoxy, CH₃ OCH₂ O--, C₁ -C₁₂ linear or branched alkanoyloxy,bromoacetoxy, benzoyloxy, azidobenzoyloxy, 3,5-(CH₃)₂ --C₆ H₃ COO--,methanesulfonyloxy, toluenesulfonyloxy, dansyloxy,(tetrahydro-2H-pyran-2-yl)oxy, or (C₁ -C₆ linear or branched alkyl)_(n)(phenyl)_(3-n) silyloxy, wherein n is 0-3.

P₃ is a radical of the formula: ##STR6## in the form of an individualisomer, an isomer mixture, a racemate or optical antipode, or apharmaceutically acceptable salt thereof, wherein L¹, L² and L³ areindividually selected from nitrogen or substituted or unsubstitutedcarbon; D¹ and D² each may be a bond or a substituted or unsubstitutedcarbon atom; D¹ may be linked to L¹, L² or both L¹ and L² to formadditional fused carbocyclic and/or heterocyclic substituted orunsubstituted rings; and D² may be linked to L², L³ or both L² and L³ toform additional fused carbocyclic and/or heterocyclic substituted orunsubstituted rings; D³ and D⁴ each are heteroatom-containing functionalgroups, the heteroatoms being selected from oxygen, nitrogen, silicon,phosphorus and sulfur; D³ may be linked to L¹, L² or both L¹ and L² toform additional fused substituted or unsubstituted carbocyclic and/orheterocyclic rings; and D⁴ may be linked to L², L³ or both L² and L³ toform additional fused substituted or unsubstituted carbocyclic and/orheterocyclic rings; provided that D¹ and D³ taken together and D² and D⁴taken together both embody at least one oxygen, nitrogen, silicon,phosphorus or sulfur atom separated from the aromatic nucleus by zero orone intervening carbon atom; and L⁴ and L⁵ are groups which, takentogether, contain about 2-40 carbon atoms, not more than 24 halogenatoms, and not more than 6 heteroatoms selected from oxygen, nitrogen,silicon, phosphorus and sulfur; provided that S_(x) E₁ may not behydroxymethyl or 1-hydroxyethyl.

P₄ is a radical of the formula: ##STR7## in the form of an individualisomer, an isomer mixture, a racemate or optical antipode, or apharmaceutically acceptable salt thereof, wherein V¹ is a bond or is acarbon atom carrying substituents individually selected from hydrogen,methyl, and halogen; V² and V³ are individually selected from oxygen,sulfur, sulfoxide, and --NV⁴ -- in which V⁴ is hydrogen or a hydrocarbonradical containing not more than 30 carbon atoms; F¹ and F²independently are groups which, taken together, contain 10-40 carbonatoms, not more than 24 halogen atoms, and not more than 8 heteroatomsselected from oxygen, nitrogen, silicon, phosphorus and sulfur, andwherein F¹ and F² may optionally be linked to form a structurecomprising 1-3 rings; F³ is hydrogen or a substituent selected frommethyl, ethyl, trifluoromethyl, 2,2,2-trifluoroethyl, cyano, vinyl,ethynyl, allyl, and propargyl; F⁴ and F⁵ each may be hydrogen or may behydrocarbon or halogenated hydrocarbon radicals which, taken together,contain not more than 40 carbon atoms, not more than 24 halogen atomsand not more than 8 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur, and wherein F⁴ and F⁵ may optionally be linked toform a structure comprising 1-3 rings; provided that V⁴, F¹, F², F⁴ andF⁵ taken together contain not more than 50 carbon atoms, not more than30 halogens and not more than 12 heteroatoms selected from oxygen,nitrogen, sulfur, silicon and phosphorus; and provided that S_(x) E₁ maynot be hydrogen, methyl, chloromethyl, hydroxymethyl, mercaptomethyl,unsubstituted carboxamido, 1-hydroxyethyl or alkanoyloxymethylene.

P₅ is a radical of the formula: ##STR8## in the form of an individualisomer, an isomer mixture, a racemate or optical antipode, or apharmaceutically acceptable salt thereof, wherein U¹ and U²,independently, are selected from hydrogen, azide, halogen, hydroxy, C₁₋₇alkoxy, C₁₋₇ alkenoxy, C₁₋₇ alkynoxy, thiol, C₁₋₇ alkanoyl, C₁₋₇saturated or unsaturated alkyl, and cyano; or U¹ and U² taken togethermay be an oxygen atom forming an epoxy group or may be an additionalbond forming an unsaturated linkage; U³ is selected from hydrogen,halogen, C₁₋₁₂ alkyl, C₁ -C₁₂ alkenyl, C₁ -C₁₂ alkynyl, C₁ -C₁₂ alkoxy,C₁ -C₁₂ alkenoxy, C₁ -C₁₂ alkynoxy, and aryl or C₇ -C₁₂ aralkyl whereinthe aryl group may be substituted by nitro, halogen, cyano, and/ordi(lower-alkyl)amino groups; U⁴ -U⁶, independently, are selected fromhydrogen, halogen, cyano, nitro, amino, di(lower-alkyl)amino, C₁₋₇saturated or unsaturated alkyl, hydroxy, C₁₋₇ saturated or unsaturatedalkoxy, C₁₋₇ carboalkoxy, C₁₋₇ alkanoyloxy, and azide; and U⁷ isselected from hydrogen, C₁₋₇ saturated or unsaturated alkyl, and C₁₋₇saturated or unsaturated alkanoyl; provided that if S_(x) E₁ ishydroxymethyl, 1-hydroxyethyl or acetoxymethyl, then S_(x) E₁ may not bebonded to C₂₉.

P₆ is a radical of the formula: ##STR9## in the form of an individualisomer, an isomer mixture, a racemate or optical antipode, or apharmaceutically acceptable salt thereof, wherein W¹ is selected fromhydrogen, halogen, hydroxy, C₁₋₅ alkoxy, C₁₋₅ alkanoyloxy and cyano; W²is selected from oxo, hydrogen, hydroxy, cyano, C₁₋₇ alkyl, C₁₋₇ alkoxy,C₁₋₇ alkanoyloxy, and halogen; W³ -W⁵ each may be hydrogen or a groupcontaining not more than 30 carbon atoms, not more than 24 halogenatoms, and not more than 8 heteroatoms selected from oxygen, nitrogen,and sulfur, and W⁴ and W⁵ taken together may form an additionalcarbocyclic or heterocyclic ring; W⁶ is hydrogen or a group containingnot more than 15 carbon atoms, not more than 12 halogen atoms and notmore than 5 heteroatoms selected from oxygen, nitrogen, and sulfur; andW⁶, taken together with W⁴ and W⁵, may complete an additionalcarbocyclic or heterocyclic ring; provided that if S_(x) E₁ ishydroxymethyl or 1-hydroxyethyl then S_(x) E₁ may not be bonded to C₂₅.

S_(x) may represent any of a broad range of connecting chains or groupsof atoms, designated S_(B), S₁, S₂, S₃, S₄, S₅ and S₆. Surprisingly,these organic functional groups may be hydrophobic in nature, with fewif any polar or heteroatoms present, may be extensivelyhalogen-substituted, or may contain one or several polar atoms such asoxygen, nitrogen, silicon, phosphorus, arsenic, boron, selenium and/orsulfur in any of numerous chemical groupings. Such functional groupingsmay even bear positive or negative charges at physiologic pH, and thevalues which are permissible for S_(x) also may include combinations ofhydrophobic, halogenated, hydrophilic and/or charged functional groups.The resultant compounds in any case generally display, variously, theprotein kinase C-modulatory, non-toxic agonist, and/or antagonisticproperties, selectivities and utilities described in this invention.

In a preferred mode of this invention, S_(B) --S₆ may comprise thefollowing values.

S_(B) is a single or double bond.

S₁ is a chain of atoms of the formula: ##STR10## wherein a, b, d, e, andg may independently be from 0 to 3; c and f may independently be 0 or 1;the sum of (a+b+c+d+e+f+g) is at least 1 but not more than 12; and if cand fare both 1, then the sum of (d+e) must be at least 1; and X and X'are as defined below.

S₂ is a chain of atoms of the formula: ##STR11## wherein h, i, k, m, p,and q may be independently be from 0 to 3; j and n may independently be0 or 1; if j and n are both 1 and 1 is 0, then the sum of (k+m) must beat least 1; if n is 1 and o is 0, then the sum of (p+q) must be at least1; the sum of (1+o) is 1-3; and the sum of (h+i+j+k+21+m+n+2o+p+q) is atleast 1 but not more than 12; and X, X', Y and Y' are as defined below.

S₃ is a chain of atoms of the formula: ##STR12## wherein r, s, u, y, a',and b' may independently be from 0 to 3; the sum of (t+z) is 0 or 1; thesum of(v+w+x) is 1; the sum of(y+z+a'+b') is at least 1; and the sum of(r+s+2t+u+2v+3w+4x+y+2z+a'+b') is at least 1 but not more than 12; andY, Y', Z¹, Z², and Z³ are as defined below.

S₄ is a chain of atoms defined by: ##STR13## wherein c', d', e', h', andi' may independently be from 0 to 3; the sum of (f+g') must be 1 or 2; fand g' may independently be 0 or 1; and the sum of(c'+d'+e'+f+g'+h'+i')is at least 1 but not more than 12; and M, M', and R^(Q) are as definedbelow.

S₅ is a chain of atoms defined by: ##STR14## wherein j', k', m', q', ands' may independently be from 0 to 3; l' and r' may each be 0 or 1, butthe sum of (l'+r') must be 1 or 2; n' and p' may each be 0 or 1, but thesum of (n'+p') must be 0 or 1; the value of o' may be 0-2; if the sum of(n'+p') is 1 and l' is 0, then q' must be at least 1; if the sumof(n'+p') is 1 and r' is 0, then m' must be at least 1; and the sum of(j'+k'+l'+m'+n'+o'+p'+q'+r'+s') is at least 1 but not more than 12; andQ, Q', X, X', and Y are as defined below.

S₆ is a chain of atoms defined by: ##STR15## wherein u', v', w', x', y',z', and m" may each be 0 or 1; t' and a" may each independently be 0-6;the sum of (t'+u'+v'+2w'+x'+2y'+z'+a") must be 0-8; b", d", e", f", h",j", k" and n" may each independently be 0 or 1; c", g", i", and l" mayeach independently be 0-3; if d" and j" are both 1, then the sumof(g"+i") must be at least 1; if either j" or k" is 1, then l" must beat least 1; if b" is 1, then the sum of(c"+g"+h"+i"+l") must be at least1; if d" is 1, then the sum of (g"+h"+i"+l") must be at least 1; and thesum of(t'+u'+v'+2w'+x'+2y'+z'+a"+b"+c"+d"+e"+2f"+g"+2h"+i"+j"+k"+l")must be 0-14; if m" is zero, R₆ ³ or R₆ ⁴ may optionally comprise anadditional bond to G² as defined below, thus completing an unsaturatedlinkage; if n" and b" are zero, R₆ ⁵ or R₆ ⁶ may optionally comprise anadditional bond to G², thus completing an unsaturated linkage; one ofthe substituents R₆ ¹ -R₆ ⁴ and/or one of the substituents R₆ ⁵ -R₆ ¹²may optionally comprise the same or different values of G¹, as definedbelow; and M, M', R^(Q), R^(Q'), R^(Q"), R^(Q'"), X, X', X", Y, Y', Z⁴,Z^(4'), Z⁵ and Z^(5') are as defined below; and wherein, for S₁ -S₆, R₁¹ through R₆ ¹² may be the same or different and each may be hydrogen,halogen or an acyclic substituent containing not more than 20 carbonatoms, not more than 16 halogen atoms, and not more than 6 heteroatomsselected from oxygen, nitrogen, sulfur, silicon and phosphorus; onesubstituent selected from R₁ ¹, R₁ ², R₂ ¹, R₂ ², R₃ ¹, R² ₃, R¹ ₄, R²₄, R¹ ₅, R² ₅, R¹ ₆ and R² ₆ may optionally comprise an additional bondcompleting an unsaturated linkage to P_(x) ; one or two of thesubstituents R₁ ¹ -R₅ ¹² may optionally comprise the same or differentvalues of G¹, as defined below; one substituent selected from R₁ ¹¹, R₁¹², R₂ ¹¹, R₂ ¹², R₃ ¹¹, R₃ ¹², R¹¹ ₄, R¹² ₄, R¹¹ ₅, R¹² ₅, R¹¹ ₆ andR¹² ₆ may optionally comprise an additional bond to E₁, therebycompleting an unsaturated linkage; one of the substituents R₁ ¹ -R₆ ¹²may be linked to either the atom in P_(x) that carries the S_(x) chainor to an atom in P_(x) adjacent thereto, to form a saturated,unsaturated or aromatic, carbocyclic or heterocyclic 4-8 membered ringoptionally containing 1-4 identical or different ring hetero membersselected from X and ═N--, the ring being optionally substituted by 1-8identical or different substituents, preferably selected from halogen,hydroxy, methoxy, ethoxy, methyl, ethyl, cyano, azide, nitro,hydroxymethyl, 1-hydroxyethyl, 2-hydroxy-2-propyl, CF₃, OCF₃, SH, SCH₃,SOCH₃, SCF₃, COOH, COOCH₃, COCH₃, CH═O, acetoxy, amino, mono- ordialkylamino totalling 1-4 carbon atoms inclusive, acetamido,N-methylacetamido, carboxamido, N-alkylated carboxamido containing 1-4carbon atoms inclusive, hydroxyacetyl and hydroxyacetoxy; provided that,for any given S₁, S₂, S₃, S₄, S₅ or S₆, but excluding P_(x) and E₁ : thetotal of carbon atoms is 25 or less; the total of halogen atoms is 16 orless; the total of oxygen atoms is 6 or less; the total of nitrogenatoms is 4 or less; the sulfur, silicon and phosphorus atoms each total3 or less; and the total of oxygen, nitrogen, silicon, phosphorus andsulfur atoms together is 8 or less.

In a preferred mode of this invention, the oxygen, nitrogen, sulfur,silicon and/or phosphorus atoms in R₁ ¹ -R₆ ¹² may be situated in avariety of functional groups such as hydroxy, amino, hydroxylamine,tertiary amine oxide, Schiff's base, hydrazine, thiol, nitro, nitroso,oxime, azide, ether, acetal, ketal, thioether, aldehyde, keto,hydrazone, carboxy, mercaptocarbonyl, mercaptothionocarbonyl, sulfonate,sulfonyl, sulfoxide, phosphate, phosphonate, phosphate ester,phosphonate ester, phosphine, phosphine oxide, thionophosphine,phosphite, phosphonium, phosphorothioate, thionophosphate ester,thiophosphonate, thionophosphonate ester, silane, silanol, silanediol,fluorinated silane, ester, amide, cyano, hydrazide, carbonate,carbamate, urea, isourea, carboxamidine, imidate, guanidine, thioester,thioamide, thiocarbonate, dithiocarbonate, thiocarbamate,dithiocarbamate, thiourea, isothiourea, thioimidate, nitroguanidine,cyanoguanidine and xanthate. Preferably, for any given S_(x), the totalof --OH groups is 3 or less, the total of --NH₂ groups is 2 or less, thetotal of --SH groups is 2 or less, and the total of --OH, --SH and --NH₂groups is 4 or less.

X, X', X" may be the same or different and are selected from: ##STR16##wherein R_(X) ¹, R_(X) ², R_(X) ¹¹ and R_(X) ¹² may independently behydrogen; R_(X) ¹ through R_(X) ¹² may be the same or different and eachmay be an acyclic substituent containing 1-20 carbon atoms, not morethan 16 halogen atoms, and not more than 6 heteroatoms selected fromoxygen, nitrogen, and sulfur, such that for any substituent the oxygenatoms total 4 or less, the nitrogen atoms total 4 or less, and thesulfur atoms total 2 or less; R_(X) ⁴, R_(X) ⁵, R_(X) ¹¹ and R_(X) ¹²may independently be hydroxy; Q and Q' are as defined below; R_(X) ¹ mayoptionally represent an additional bond to P_(x), thus completing anunsaturated linkage; one to four of the substituents R_(X) ¹ -R^(X) ¹²may optionally comprise the same or different values of G¹, as definedbelow.

Y and Y' may be the same or different and are selected from: ##STR17##wherein R_(Y) ¹ and R_(Y) ², and R_(Y) ³ and R_(Y) ⁴, each pair beingcis or trans relative to one another, may be the same or different andeach may be hydrogen or an acyclic substituent containing not more than20 carbon atoms, not more than 16 halogen atoms, and not more than 6heteroatoms selected from oxygen, nitrogen, and sulfur, such that forany substituent the oxygen atoms total 4 of less, the nitrogen atomstotal 4 or less, and the sulfur atoms total 2 or less; R_(Y) ¹ and R_(Y)² may also independently be halogen; one or two of the substituentsR_(Y) ¹ -R_(Y) ⁴ may optionally comprise the same or different values ofG¹, as defined below; and one of the substituents R_(X) ¹ -R_(X) ¹² andR_(Y) ¹ -R_(Y) ⁴ may be linked to either the atom in P_(x) that carriesthe chain containing X, X', and/or X" or to an atom in P_(x) adjacentthereto, to form a saturated, unsaturated or aromatic, carbocyclic orheterocyclic 4-8 membered ring defined as for the analogous R₁ ¹ -R₆ ¹²-containing ring above.

In a preferred embodiment, the substituents R_(X) ¹ -R_(X) ¹² and R_(Y)¹ -R_(Y) ⁴ are selected from hydroxy, amino, thiol, nitro, azide, ether,thioether, aldehyde, keto, carboxy, mercaptocarbonyl,mercaptothionocarbonyl, sulfonate, sulfonyl, sulfoxide, ester, amide,cyano, carbonate, carbamate, urea, isourea, carboxamidine, guanidine,thioester, thioamide, thiourea, nitroguanidine, cyanoguanidine andxanthate.

Z¹ is selected from: ##STR18##

Z² is selected from: ##STR19##

Z³ is selected from: ##STR20##

Z⁴ and Z^(4') independently may be: ##STR21##

Z⁵ and Z^(5') independently may be: ##STR22## wherein the substituentsR_(Z) ¹ -R_(Z) ³⁰ are generally selected from hydrogen, halogen in caseswhere a chemically stable structure results, and a radical containingabout 1-12 carbon atoms and optionally containing 0-12 halogens and 0-6heteroatoms selected from oxygen, nitrogen and sulfur.

In a preferred embodiment of the invention, the substituents R_(Z) ¹-R_(Z) ³⁰ comprise a range of saturated or unsaturated substituents asdescribed below, wherein the terms alkyl, halogenated alkyl and acyl aretaken to include alkenyl, alkynyl, alkenoyl and alkynoyl and theirhalogenated forms.

Thus: R_(Z) ²⁶ be any of the values specified for R_(X) ¹ -R_(X) ¹²above; R_(Z) ¹⁸ and R_(Z) ¹⁹ individually may be --O--, --S--, or--NR_(Z) ²¹ --, wherein R_(Z) ²¹ may be hydrogen, C₁₋₄ alkyl,2-hydroxyethyl, 2-hydroxy-n-propyl, 2-acetoxyethyl, or2-acetoxy-n-propyl; R_(Z) ¹⁷ and R_(Z) ²⁰ individually may be hydrogenor a substituent selected from C₁₋₄ alkyl; C₁₋₄ alkoxy; C₁₋₄ alkylthio;phenoxy or thiophenoxy optionally substituted by methyl, hydroxy,hydroxymethyl, thiol, carboxy, carboxymethyl, amino, methoxy, halogen,and/or nitro; or amino optionally mono- or disubstituted by C₁₋₄ alkylor monosubstituted by cyano, nitro or phenyl optionally substituted byhalogen, hydroxy, hydroxymethyl, thiol, carboxy, carboxymethyl, amino,and/or nitro; R_(Z) ¹ -R_(Z) ¹⁶, R_(Z) ²² -R_(Z) ²⁵ and R_(Z) ²⁷ -R³⁰_(Z) may be hydrogen, a saturated or unsaturated substituent selectedfrom C₁₋₆ alkyl, C₁₋₆ halogenated alkyl, and cyclohexyl, or may bephenyl or benzyl, each optionally substituted by methyl, ethyl, hydroxy,hydroxymethyl, thiol, carboxy, carboxymethyl, amino, methoxy, nitro,cyano, trifluoromethyl, and/or halogen; R_(Z) ¹ -R_(Z) ⁴, R_(Z) ⁷, R_(Z)¹⁰, R_(Z) ¹², R¹³ _(Z) and R²⁷ _(Z) -R³⁰ _(Z) may also be independentlyselected from C₁₋₆ acyl, C₁₋₆ halogenated acyl, C₂₋₆ monohydroxyacyl,and C₂₋₆ hydroxyalkyl; R_(Z) ⁵ and R_(Z) ⁶ may also be independentlyselected from C₁₋₆ hydroxyalkyl, C₁₋₆ alkoxy, C₁₋₆ hydroxyalkoxy; andR_(Z) ⁸, R_(Z) ⁹, R_(Z) ²³ and R_(Z) ²⁴ may also independently be C₂₋₆hydroxyalkyl; one of the substituents R_(Z) ¹ -R_(Z) ¹⁷, R_(Z) ²⁰, R^(Z)²¹, R_(Z) ²² and R²⁵ _(Z) may be linked to either the atom in P_(x) thatcaries the chain containing Z¹ or to an atom in P_(x) adjacent thereto,to form a saturated, unsaturated, or aromatic, carbocyclic orheterocyclic 4-8 membered ring optionally containing 1-4 other identicalor different hetero ring members selected from O, S, ═N--, and NH, thering being optionally substituted on its carbon and/or NH members by 1-8identical or different substituents selected from halogen, hydroxy,methoxy, ethoxy, methyl, ethyl, cyano, azide, nitro, hydroxymethyl,1-hydroxyethyl, 2-hydroxy-2-propyl, CF₃, OCF₃, SH, SCH₃, SOCH₃, SCF₃,COOH, COOCH₃, COCH₃, CH═O, acetoxy, amino, mono- or dialkylaminototalling 1-4 carbon atoms inclusive, acetamido, N-methylacetamido,carboxamido, N-alkylated carboxamido containing 1-4 carbon atomsinclusive, hydroxyacetyl and hydroxyacetoxy; R_(Z) ²⁶ may comprise G¹ asdefined below; an optional ring may be formed between R_(Z) ²⁶ and P_(x)as described above for R_(X) ¹ -R_(X) ¹² ; R_(Z) ¹, R_(Z) ⁴, R⁷ _(Z),R²⁶ _(Z) or R_(Z) ²⁷ may comprise an additional bond to P_(x), thuscompleting an unsaturated linkage; and only one of the substituentsR_(Z) ¹ -R_(Z) ²⁵ may be substituted or unsubstituted phenyl or benzyl.

M and M' independently may be: ##STR23## wherein R_(M) ¹ and R_(M) ² maybe the same or different and each may be hydrogen or a saturated orsingly or multiply unsaturated, straight or branched, acyclicsubstituent containing 1-20 carbon atoms, not more than 16 halogenatoms, and not more than 6 heteroatoms selected from oxygen, nitrogen,and sulfur, in which the oxygen atoms total 4 or less, the nitrogenatoms total 4 or less, and the sulfur atoms total 2 or less, theheteroatoms being preferably situated in functional groups selected fromhydroxy, amino, thiol, nitro, azide, ether, thioether, aldehyde, keto,carboxy, ester, amide, cyano, nitroguanidine, and cyanoguanidine; R_(M)¹ may optionally comprise an additional bond to P_(x) group, thuscompleting an unsaturated linkage; R_(M) ¹ or R_(M) ² may optionallycomprise the same or different values of G¹, as defined below; R_(M) ¹or R_(M) ² may be linked to either the atom in P_(x) that carries thechain containing M and/or M' or to an atom in P_(x) adjacent thereto, toform a saturated, unsaturated or aromatic, carbocyclic or heterocyclic4-8 membered ring defined as for the analogous R₁ ¹ -R₆ ¹² -containingring above.

Q-Q'" independently may be:

    ═O═S═N--R.sub.Q.sup.1 or ═N--O--R.sub.Q.sup.2

wherein R_(Q) ¹ and R_(Q) ² may be the same or different and each mayhave the values specified above for R_(M) ¹ and R_(M) ² ; R_(Q) ¹ and/orR_(Q) ² may optionally comprise the same or different values of G¹, asdefined below; R_(Q) ¹ may be linked to either the atom in P_(x) thatcarries the chain containing Q and/or Q' or to an atom in P_(x) adjacentthereto, to form a saturated, unsaturated or aromatic, carbocyclic orheterocyclic 4-8 membered ring defined as for the analogous R₁ ¹ -R₆ ¹²-containing ring above.

R^(Q) -R^(Q'") are independently selected from: ##STR24## wherein R_(Q)³ and R_(Q) ⁴ may be the same or different and each may be selected fromhalogen and the values specified above for R_(M) ¹ and R_(M) ² ; R_(Q) ³and/or R_(Q) ⁴ may optionally comprise the same or different values ofG¹, as defined below; Q and Q' are as defined above; one of R_(Q) ³ andR_(Q) ⁴ may be linked to either the atom in P_(x) bonded to the chainthat carries R^(Q) or to an atom in P_(x) adjacent thereto, to form asaturated, unsaturated or aromatic, carbocyclic or heterocyclic 4-8membered ring defined as for the analogous R₁ ¹ -R₆ ¹² -containing ringabove.

G¹ and G² independently comprise a group containing 1-3 fused orseparate, saturated, unsaturated or aromatic, carbocyclic orheterocyclic 4-8 membered rings, each ring optionally containing 1-4identical or different hetero ring members selected from X and ═N--,each ring being optionally substituted on its carbon and/or NH membersby 1-8 identical or different substituents, preferably selected fromhalogen, hydroxy, methoxy, ethoxy, methyl, ethyl, cyano, azide, nitro,hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 2-hydroxy-2-propyl, CF₃,OCF₃, SH, SCH₃, SOCH₃, SCF₃, COOH, COOCH₃, COCH₃, CH═O, acetoxy, amino,mono- or dialkylamino totalling 1-4 carbon atoms inclusive, acetamido,N-methylacetamido, carboxamido, N-alkylated carboxamido containing 1-4carbon atoms inclusive, hydroxyacetyl and hydroxyacetoxy;

wherein for G¹ the optional second and third rings may be fused to thefirst ring and/or to one another or may be separate rings connected toone another and/or to the atom bearing G¹ by a single or double bond orby an intervening substituted or unsubstituted, linear or branched,saturated or unsaturated chain containing not more than 8 carbon atoms,not more than 8 halogens, and not more than 4 heteroatoms selected fromoxygen, nitrogen, silicon, phosphorus and sulfur;

and wherein for G² the first ring is singly or doubly bonded to P_(x) orto a component atom of the S₆ chain connecting P_(x) and G², and theoptional second and third rings may be fused to the first ring or to oneanother or may be separate rings connected to one another and/or to thefirst ring by single or double bonds or by an intervening substituted orunsubstituted, linear or branched, saturated or unsaturated chaincontaining not more than 8 carbon atoms, not more than 8 halogens, andnot more than 4 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur.

The capping group E_(o) that terminates the connecting chain also may beselected from any of a surprisingly broad array of chemical groupings,and these chemical groupings can be composed of a far larger number ofatoms than is found in the hydroxymethyl or 1-hydroxyethyl group. Thesechemical groupings may include, without limitation, hydrophobic entitiessuch as alkyl, hydrogen, and halogenated alkyl, or may include, withoutlimitation, quite hydrophilic organic functional groups, such ashydroxy, thiol, carboxy and carboxy esters, amines, etc. It iswell-known in the art that organic functional groups spanning a widerange of properties, from ionized and very hydrophilic to veryhydrophobic, can be formed from multi-atom groupings of elementsselected from carbon, hydrogen, halogen, oxygen, nitrogen, silicon,phosphorus, arsenic, boron and selenium. Indeed, for this invention thesingle restriction appears to be that S_(o) E_(o) taken together shouldnot be hydroxymethyl or 1-hydroxyethyl bonded in the usual position inthe parent compounds, since such compounds correspond to theskin-inflammatory and often tumor-promoting parent natural products.

Thus, E_(o) may comprise E₁, wherein E₁ is selected from ═O, ═S, ═NH,═NOR_(E) ⁸ wherein R_(E) ⁸ is hydrogen or a C₁ -C₈ normal or branchedalkyl radical, ═N--NH₂, hydrogen, halogen, --OH, --SH, --NH₂, --NH--NH₂,--N₃, --CN, --NO, --NO₂, --NHOH, --ONH₂, or is selected from: ##STR25##wherein T¹ is selected from --O--, --S--, and --NH--; T² is selectedfrom ═O, ═S, and ═N--R_(E) ⁶, in which R_(E) ⁶ may be hydrogen, hydroxy,cyano, or nitro; T^(2') is selected from ═O and ═S; T³, T⁴ and T^(4')are independently selected from --OH, --NH₂, --SH, --N₃, --NH--NH₂, and--NH--OR_(E) ⁷, in which R_(E) ⁷ may be hydrogen, C₁₋₃ alkyl or C₁₋₃acyl; T³ may also be hydrogen or halogen; T⁵ -T^(5") are independentlyselected from hydrogen and hydroxy; T⁵ may also be halogen; R_(E) ¹, isselected from hydrogen, halogen, hydroxy, nitro, nitroso, cyano, azide,--NH₂, --NH--OH, --SH, --O--NH₂, --NH--NH₂, --T¹ --C(═T²)--T³,--C(═T²)--T³, --SiT⁵ T^(5') T^(5"), --T¹ --S(═O)(═T²)--T⁴,--S(═O)(═T²)--T⁴, --T¹ --P(--T⁴)--T^(4') ; --P(--T⁴)--T^(4'), --T¹--P(═T^(2'))(--T³)--T⁴, and --P(═T^(2'))(--T³)--T⁴ ; R_(E) ² and R_(E) ³are individually selected from hydrogen, --C(═T²)--T³, cyano, nitro,azide, halogen and a C₁ -C₁₅ straight or branched chain, saturated,unsaturated and/or aromatic substituent optionally containing not morethan 10 halogen atoms and not more than 4 heteroatoms selected fromoxygen, nitrogen and sulfur.

Finally, if R_(E) ¹ is cyano or --C(═T²)--T³, then R_(E) ² or R_(E) ³may optionally be selected from --SiT⁵ T^(5') T^(5"), --T¹--P(═T^(2'))(--T³)--T⁴, and --P(═T^(2'))(--T³)--T⁴ ; and R_(E) ⁴ andR_(E) ⁵ are individually selected from hydrogen, halogen, cyano, nitro,--C(═T²)--T³, --T¹ --C(═T²)--T³, --CR_(E) ¹ R_(E) ² R_(E) ³, --SiT⁵T^(5') T^(4"), --S(═O)(═T²)--T⁴, and --P(═T^(2'))(--T³)--T⁴.

In this invention, novel phorboids of the P₁ diterpene class are furtherillustrated by the general structure P_(1R), ##STR26## wherein carbons(1 and 2) or (2 and 3) may be joined by a double bond; carbons (5 and 6)or (6 and 7) may be joined by a double bond; S₁ E₁ may be bonded tocarbon 5, 6 or 7; R_(A1) represents not more than 8 identical ordifferent substituents bonded independently via single and/or doublebonds to carbons 1, 2 and/or 3, which substituents may independently behalogen(s) and/or other groups, which halogens and groups taken togethercontain a total of not more than 30 carbon atoms, not more than 24halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur; R_(A3) represents not morethan 6 identical or different substituents bonded independently viasingle and/or double bonds to carbons 5, 6 and/or 7, which substituentsmay independently be halogen(s) and/or other groups, which halogens andgroups taken together contain not more than 12 carbon atoms, not morethan 8 halogen atoms, and not more than 5 heteroatoms selected fromoxygen, nitrogen, silicon, phosphorus and sulfur; and R_(A4) representsnot more than 10 identical or different substituents bondedindependently via single or double bonds to carbons 9, 11, 12, 13 and/or14, which substituents may independently be halogen(s) and/or othergroups, the group or groups optionally completing 1-3 additional ringsthrough bonds among themselves and/or 1-5 additional rings when takentogether with the 5-membered ring and its substituent(s) R_(A1), whichhalogen and groups taken together, include not more than 50 carbonatoms, not more than 24 halogen atoms, and not more than 15 heteroatomsselected from oxygen, nitrogen, silicon, phosphorus and sulfur.

Readily available embodiments of P_(1R) are illustrated by, but notlimited by, structures carrying a substituted or unsubstitutedcyclopropyl ring, forming P_(1P) ##STR27## wherein the R_(A5) and R_(A6)radicals may independently be hydrogen, halogen and/or other groups,which halogens and groups taken together contain a total of not morethan 30 carbon atoms, not more than 24 halogen atoms and not more than 9heteroatoms selected from oxygen, nitrogen, silicon, phosphorus andsulfur.

In compounds further illustrating P_(1P), carbon 2 carries a methylgroup, J¹ is hydroxy, carbon 9 carries a hydroxy group in the alphaconfiguration, carbons 10 and 14 carry alpha hydrogens, carbon 11carries a methyl group in the alpha configuration and R_(A5) and R_(A6)are both methyl, forming a preferred parent structure for use in thepresent invention, P_(1PP) : ##STR28##

In another embodiment of P_(1R), an orthoester structure is bonded tothe 6-membered ring via one orthoester oxygen atom in the alphaconfiguration to carbon 9 and via two orthoester oxygen atoms in thealpha configuration to any two of carbons 12-14, forming P_(1RR) :##STR29## wherein R_(A7) represents hydrogen, halogen or other group,the group optionally completing 1 additional ring to carbon 1, whichgroup includes not more than 30 carbon atoms, not more than 16 halogenatoms, and not more than 9 heteroatoms selected from oxygen, nitrogen,silicon, phosphorus and sulfur.

In a preferred embodiment of P_(1RR), carbon 2 carries a methyl group,J¹ is hydroxy, carbon 10 carries a hydrogen atom in the alphaconfiguration, carbon 11 carries a methyl group in the alphaconfiguration and carbon 13 carries an isopropenyl group, formingP_(1RRR) : ##STR30##

Novel phorboids of the P₁ diterpene class may also advantageously embodythe general structure illustrated by formula P_(1I) : ##STR31## whereinR_(A2I) represents not more than 8 identical or different substituentsbonded independently via single and/or double bonds to carbons 1, 2and/or 3, which substituents may independently be halogen(s) and/orother groups, which halogens and groups taken together contain a totalof not more than 30 carbon atoms, not more than 24 halogen atoms and notmore than 9 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur; R_(A3I) represents not more than 6 identical ordifferent substituents bonded independently via single and/or doublebonds to carbons 5, 6 and/or 7, which substituents may independently behalogen(s) and/or other groups, which halogens and groups taken togethercontain not more than 12 carbon atoms, not more than 8 halogen atoms,and not more than 5 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur; and R_(A4I) represents not more than 10 identicalor different substituents bonded independently via single or doublebonds to carbons 9, 11, 12, 13 and/or 14, which substituents mayindependently be halogen(s) and/or other groups, the group or groupsoptionally completing 1-3 additional rings through bonds amongthemselves and/or 1-5 additional rings when taken together with the5-membered ring and its substituent(s) R_(A2I), which halogen and groupstaken together, include not more than 50 carbon atoms, not more than 24halogen atoms, and not more than 15 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur.

In compounds further illustrating P_(1I), carbons 13 and 14 of carry asubstituted or unsubstituted cyclopropyl ring, thus forming P_(1IN) :##STR32## wherein the R_(A5I) and R_(A6I) radicals may independently behydrogen, halogen and/or other groups, which halogens and groups takentogether contain a total of not more than 30 carbon atoms, not more than24 halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur.

In preferred embodiments of P_(1IN) as a parent structure for thepractice of the present invention, carbon 2 carries a methyl group, J¹is hydroxy, carbon 11 carries a methyl group in the alpha configuration,carbon 14 carries an alpha hydrogen and R_(A5I) and R_(A6I) are bothmethyl, forming P_(1INL) : ##STR33##

Among very diverse parent phorboids of the P₂ class ofindole/indene/benzofuran/benzothiophene derivatives, the novel compoundsof this invention may advantageously embody the general structureP_(2NN), ##STR34## wherein K_(B1) represents 1-3 identical or differentsubstituents located independently at carbons 5, 6, and/or 7, whichsubstituents may independently be hydrogen, halogen and/or other groupswhich, taken together, contain not more than 40 carbon atoms, not morethan 24 halogen atoms, and not more than 9 heteroatoms selected fromoxygen, nitrogen, silicon, phosphorus, and sulfur, the groups beingoptionally connected to one another, to K⁸ and/or to K_(B2) to form 1-3additional carbocyclic or heterocyclic rings; and wherein K_(B2) is,hydrogen or a group which contains not more than 20 carbon atoms, notmore than 24 halogen atoms, and not more than 6 heteroatoms selectedfrom oxygen, nitrogen, silicon, phosphorus and sulfur, this group beingoptionally connected to K_(B1) or K₁ to form an additional carbocyclicor heterocyclic ring.

In compounds further illustrating P_(2NN), K¹ and K³ are hydrogen and K²is --NH--, forming P_(2NNN) ##STR35## and K⁴ is hydrogen or other groupcontaining not more than 20 carbon atoms, not more than 24 halogenatoms, and not more than 9 heteroatoms selected from oxygen, nitrogen,silicon, phosphorus and sulfur, the group being optionally connected toK⁸ to form an additional ring.

Two particularly preferred embodiments of P_(2NNN) comprise P_(2L) andP_(2T), wherein P_(2L) is ##STR36## wherein K_(B1), is hydrogen, halogenor other group which contains not more than 40 carbon atoms, not morethan 24 halogen atoms, and not more than 9 heteroatoms selected fromoxygen, nitrogen, silicon, phosphorus and sulfur, this group beingoptionally connected to K_(B2) to form an additional ring; and whereinP_(2T) is ##STR37## wherein a four-carbon saturated, unsaturated oraromatic ring connects positions 6 and 7 and substituents K₁₀ -K₁₃ mayindependently be absent in favor of unsaturated linkages or may behydrogen, halogen and/or other groups which, taken together, contain notmore than 36 carbon atoms, not more than 24 halogen atoms, and not morethan 9 heteroatoms selected from oxygen, nitrogen, silicon, phosphorusand sulfur.

It will be appreciated that the many different permissible changes tothe hydroxymethyl or 1-hydroxyethyl groups of the parent phorboids leadto diverse compounds with diverse biological properties, and differentembodiments will be preferred for different utilities. If differentprotein kinase C isotypes and other proteins bearing phorboid-typebinding sites have different biological functions, as has beenextensively hypothesized and to some extent demonstrated in biologicalexperiments, then the novel compounds of this invention with differingactivity on different protein kinase C isotypes will obviously display awide range of differing utilities.

For example, a preferred set of compounds for anti-viral activity,including anti-retroviral and anti-Human Immunodeficiency Virus (HIV)activities, in human cells is generated by replacing the hydroxymethylor 1-hydroxyethyl groups of parent phorboids with the following organicfunctional groups: (i) dihalomethyl, trihalomethyl, --N₃, --NH₂, --CN,--COOH, --CH═CHCOOH, --C.tbd.CCOOH, --CSSH, --COSH, --SO₂ H, --SO₃ H,--P(═O)(R_(a) ^(a))OH, --P(═S)(R_(a) ^(a))OH, --P(═O)(OR_(a) ^(a))OH,--P(═S)(OR_(a) ^(a))OH, --CH═NOH, --CH═NOCH₃, --CH═N(→O)CH₃,--C(CH₃)═NOH, --CH═CHR_(a) ^(a), --C.tbd.C--R_(a) ^(a), --CH₂C.tbd.C--R_(a) _(a), --Si(CH₃)₂ OH, --Si(OH)₂ CH₃, --Si(CH₃)₂ F,--Si(CH₃)₂ R_(a) ^(a), --CH₂ Si(CH₃)₂ R_(a) ^(a), or ═CHR_(a) ^(a), inwhich R_(a) ^(a) is hydrogen or C₁₋₁₂ linear or branched, saturated,unsaturated and/or aromatic hydrocarbon optionally substituted by notmore than 16 halogens; or (ii) --CH₂ -- or --CH(CH₃)--, to either ofwhich is bonded --R_(a) ^(a), --F, --N₃, --NH₂, --CN, --COOH, --COSH,--CSSH, --Si(CH₃)₂ OH, --Si(OH)₂ CH₃, --Si(CH₃)₂ F, --SO₂ H, --SO₃ H,--PO₃ H₂, --P(═O)(R_(a) ^(a))OH, --P(═S)(R_(a) ^(a))OH, --P(═O)(OR_(a)^(a))OH, --P(═S)(OR_(a) ^(a))OH, the o-, m- or p-isomer of --M--C₆ H₄CH₂ --T³, the o-, m- or p-isomer of --C₆ H₄ CH₂ --T³, --SR_(a) ^(a),--SCH₂ CH₂ OH, --S(═O)--CH₂ CH₂ OH, --S(CH₂)₃ OH, --S(CH₂)₄ OH, --SCH₂CH₂ SH, --M--C(═T²)--M'--R_(a) ^(a) except --OC(═O)NH₂, --OCH₂ C(═O)CH₃,--OCH₂ C(═NOH)CH₃, the o-, m- or p-isomer of --M--C(═T²)--M'--C₆ H₄--T³, or the o-, m- or p-isomer of --M--C(═T²)--M'--C₆ H₄ CH₂ --T³.

Preferred compounds for anti-viral use, for example, incorporate parentradicals selected from P_(1PP), P_(1RRR), P_(1INL), P_(2L) and P_(2T),bearing functionally diverse S_(x) E₁ groups selected from (i)dihalomethyl, trihalomethyl, --N₃, --NH₂, --CN, --COOH, --CH═CHCOOH,C═CCOOH, --CSSH, --COSH, --SO₂ H, --SO₃ H, --P(═O)(R_(a) ^(a))OH,--P(═S)(R_(a) ^(a))OH, --CH═NOH, --Si(CH₃)₂ OH, --Si(OH)₂ CH₃, or (ii)--CH₂ -- or --CH(CH₃)--, to either of which is bonded --R_(a) ^(a), --F,--N₃, --NH₂, --CN, --COOH, --COSH, --CSSH, --Si(CH₃)₂ OH, --Si(OH)₂ CH₃,--SO₃ H, --PO₃ H₂, --P(═O)(R_(a) ^(a))OH, the o-, m- or p-isomer of--M--C₆ H₄ CH₂ --T³, the o-, m- or p-isomer of --C₆ H₄ CH₂ --T³,--SR_(a) ^(a), --SCH₂ CH₂ OH, --S(═O)--CH₂ CH₂ OH, --S(CH₂)₃ OH,--S(CH₂)₄ OH, --SCH₂ CH₂ SH, the o-, m- or p-isomer of--M--C(═T²)--M'--C₆ H₄ --T³, or the o-, m- or p- isomer of--M--C(═T²)--M'--C₆ H₄ CH₂ --T³, in which R_(a) ^(a) is hydrogen orC₁₋₁₂ linear or branched, saturated, unsaturated and/or aromatichydrocarbon optionally substituted by not more than 16 halogens.

The compounds of this invention have been found to possess valuablepharmacological properties for human and veterinary medicine. Fortherapeutic use in humans or animals the compounds of this invention aredispensed in unit dosage form comprising 0.001 to 1000 mg per unitdosage in a pharmaceutically acceptable carrier. In particular, unitdosages in the range of 0.1 to 100 mg are preferred. The compounds ofthis invention may also be incorporated in topical formulations inconcentrations of about 0.001 to 10 weight percent, with concentrationsof 0.01 to 10 weight percent being preferred.

It will be appreciated that the actual preferred amounts of activecompound in a specific case will vary according to the specific compoundbeing utilized, the particular compositions formulated, the mode ofapplication, and the particular sites and organism being treated.Compounds of this invention having higher potencies should be used ingenerally smaller amounts, and compounds with lower potencies should beused in generally larger amounts. Dosages for a given host, whether asmall animal such as a cat or a human patient, can be determined usingconventional considerations, e.g., the host's weight or body surfacearea. In general, the compounds of the present invention areadministered in quantities of about 0.0001 to about 1000 mg/kg of bodyweight, and quantities of about 0.01 to about 100 mg/kg of body weightare preferred.

As specific examples, representative compounds of this inventionvariously block inflammation; show cytostatic and/or cytotoxic activityagainst very diverse types of human cancer cells representative ofseveral human cancers such as leukemia, carcinoma and melanoma; inhibitthe infection of human lymphoid cells by HIV; and induce production ofthrombolytic activity. These effects are demonstrated by (i) inhibitionin standard topical in vivo mouse ear inflammation tests whereininflammation by established agonists such as PMA and the ionophoreA23187 are blocked; (ii) by the inhibition of proliferation of humanleukemic cells in culture via induction of differentiation; (iii) byassays of cytotoxicity against human carcinoma cancer cells; (iv) byassays of inhibition of growth of human melanoma cells; (v) by assays ofinhibitory activity against acute and chronic HIV viral replication inhuman lymphocyte cells; and (vi) by measurement of stimulation offibrinolytic activity in cultured cells. These demonstrations ofefficacy are achieved at doses per ear, per kg of body weight or per kgof bodily fluid equivalent, using numerous representative compounds ofthe present invention, as follows: (i) about 0.01-1000 nanogram/ear,corresponding to about 0.0001-10 mg per square meter of body surfacebeing treated; (ii) about 0.1-100 mg/kg fluids; (iii) about 0.1-100mg/kg fluids; (iv) about 1-100 mg per kg of fluids; (v) about 0.1-100mg/kg fluids; and (vi) about 0.1-100 mg/kg fluids, respectively.

The activities of representative compounds of this invention against thethree diverse types of human cancer cells described above areparticularly noteworthy. Cancer is in fact a broad classificationcontaining at least 110 clinically different types of tumorous diseases.The activities of compounds of this invention against three verydifferent types of cancer cells demonstrate not only those utilitiesagainst specific human cancer cells but also significant breadth for theanti-cancer effects of this invention beyond a single class of cancerdisease, indicative of additional anti-cancer utilities.

The anti-viral activities of numerous compounds of this invention arealso of great importance. For example, the tests demonstrating theanti-HIV properties of these compounds [see Examples 116-119] werecarried out in widely validated and accepted cellular assays of HIVinfectivity in human cells that are indicative of in vivo activity. Thusthe anti-HIV properties of the compounds of this invention relatedirectly to the in vivo activities of standard anti-HIV reversetranscription inhibitors such as azidothymidine, dideoxyinosine,dideoxycytidine and non-nucleoside reverse transcription inhibitors,HIV-protease inhibitors and inhibitors of tat-gene function, which arefully active in the assay by which the compounds of this invention weretested. This is in contrast to many in vitro HIV-related assays that areunable to provide predictive information about the anti-HIV effects oftest compounds in living cells, such as assays of inhibition againstisolated HIV enzymes. For example, the inadequacy of isolated enzymeassays was indicated by the finding that, of two compounds able toinhibit the HIV protease when assayed on the purified protease enzyme,only one of the compounds was able to inhibit HIV infectivity in thewhole-cell assay in human lymphocytes [T. K. Antonucci et al.,"Characterizations of HIV-1 protease inhibitors" in Innovations intherapy of human viral diseases: a Wellcome Symposium, Dec. 6-9, 1992,Book of Abstracts, Page 2.]

The compounds of this invention also show selective effects asantagonists for protein kinase C in some cases, as noninflammatoryagonists for protein kinase C in other cases, and as selective ligandsfor protein kinase C and/or for phorboid receptors.

Thus, these compounds can be used as agents for the abrogation ofpathophysiological conditions and disease states in applications such asanti-inflammatory, anti-psoriatic, anti-cancer, anti-ulcer,anti-hypertensive, anti-asthma, anti-arthritic, anti-autoimmune,anti-nociceptive, anti-secretory, anti-parasitic, anti-amoebic,anti-viral including anti-HIV replication, in prophylaxis againstinfection by any HIV form, and any other application in whichpathological involvement of protein kinase C is found.

For example, evidence for involvement of protein kinase C in thephysiology of many human and animal pathogenic viruses has long beenknown, particularly from experiments in which a standard protein kinaseC activator such as a phorbol ester greatly stimulates viral productionfor many different kinds of viruses [see, for example: H. zur Hausen etal., "Persisting oncogenic herpesvirus induced by the tumour promoterTPA", Nature 272, 373-375 (1978); H. zur Hausen et al., "Tumorinitiators and promoters in the induction of Epstein-Barr virus", Proc.Natl. Acad. Sci. USA 76, 782-785 (1979); D. V. Ablashi et al.,"Increased infectivity of oncogenic Herpes viruses of primates withtumor promoter 12-O-tetradecanoylphorbol-13-acetate", Proc. Soc. Exp.Biol. Med. 164, 485-490 (1980); G. Colletta et al., "Enhancement ofviral gene expression in Friend erythroleukemia cells by12-O-tetradecanoylphorbol-13-acetate", Cancer Research 40, 3369-3373(1980); S. K. Arya, "Phorbol ester-mediated stimulation of the synthesisof mouse mammary tumour virus", Nature 284, 71-72 (1980); K. B. Hellmanand A. Hellman, "Induction of type-C retrovirus by the tumor promoterTPA", Int. J. Cancer 27, 95-99 (1981); E. Amtmann and G. Sauer,"Activation of non-expressed bovine papilloma virus genomes by tumourpromoters", Nature 296, 675-677 (1982); L. S. Kucera et al.,"12-O-Tetradecanoyl-phorbol-13-acetate enhancement of the tumorigenicpotential of Herpes Simplex virus type 2 Transformed cells", Oncology40, 357-362 (1983); and V. Wunderlich et al., "Enhancement of primateretrovirus synthesis of tumor promoters", Symposium on role ofcocarcinogens and promoters in human and experimental carcinogenesis,16-18 May, 1983, Budapest, Hungary, Book of Abstracts, p. 88].

Similar experiments indicate involvement of protein kinase C in the lifecycle of HIV, and initial molecular genetics studies helped illuminatethe mechanisms by which cellular protein kinase C can influence HIV[see, for example: S. Harada et al., "Tumor promoter, TPA, enhancesreplication of HTLV-III/LAV", Virology 154, 249-258 (1986); H. Dinter etal., "In Vitro activation of the HIV-1 enhancer in extracts from cellstreated with a phorbol ester tumor promoter", EMBO Journal 6, 4067-4071(1987); J. D. Kaufman et al., "Phorbol ester enhances humanimmunodeficiency virus-promoted gene expression and acts on a repeated10-base-pair functional enhancer element", Mol. Cell. Biol. 7, 3759-3766(1987); and M. Siekevitz et al., "Activation of the HIV-1 LTR by T cellmitogens and the trans-activator protein of HTLV-I", Science 238,1575-1578 (1987)].

These and later molecular genetics-based virological investigationsprovided clear mechanistic explanations for the effects, observed muchearlier, of protein kinase C modulators on the life cycles of numeroushuman and animal viruses. Such studies showed that many viruses containgenetic control elements, called enhancers, whose functions incontrolling viral expression involve the protein kinase C of the hostcell. Of particular importance for the role of protein kinase C invirus-cell interactions are the enhancers known as AP-1 and NF-κB [see,for example: J. E. Marich et al., "The phylogenetic relationship andcomplete nucleotide sequence of human papillomavirus Type 35", Virology186, 770-776 (1992); R. L. Smith et al., "Activation of second-messengerpathways rectivates latent Herpes Simplex virus in neuronal cultures",Virology 188, 311-318 (1992); S. L. Gdovin and J. E. Clements,"Molecular mechanisms of visna virus tat: identification of the targetsfor transcriptional activation and evidence for a post-transcriptionaleffect", Virology 188, 438-450 (1992); D. S. Shih et al., "Involvementof FOS and JUN in the activation of visna virus gene expression inmacrophages through an AP-1 site in the viral LTR", Virology 190, 84-91(1992); A. Mirza, "Stimulation of adenovirus early gene expression byphorbol ester: its possible mechanism", Virology 190, 645-653 (1992); E.J. Wade et al., "An AP-1 binding site is the predominant cis-actingregulatory element in the 1.2-kilobase early RNA promoter of humancytomegalovirus", J. Virology 66, 2407-2417 (1992); F. Stubenrauch etal., "Late promoter of human papillomavirus Type 8 and its regulation",J. Virology 66, 3485-3493 (1992); F. Thierry et al., "Two AP-1 sitesbinding JunB are essential for human papillomavirus Type 18transcription in keratinocytes", J. Virology 66, 3740-3748 (1992); J.Liu et al., "Specific NF-κB subunits act in concert with tat tostimulate human immunodeficiency virus Type 1 transcription", J.Virology 66, 3883-3887 (1992); W. A. Jensen et al., "Inhibition ofprotein kinase C results in decreased expression of bovine leukemiavirus", J. Virology 66, 4427-4433 (1992); K. Shiroki et al., "AdenovirusE1A proteins stimulate inositol phospholipid metabolism in PC12 cells",J. Virology 66, 6093-6098 (1992); J. C. cross et al., "Transactivationby hepatitis B virus X protein is promiscuous and dependent onmitogen-activated cellular serine-threonine kinases", Proc. Natl. Acad.Sci. USA 90. 8078-8082 (1993); and A. S. Kekule et al., "Hepatitis Bvirus transactivator HBx uses a tumour promoter signalling pathway",Nature 361, 742-745 (1993)].

The compounds of this invention can also be used in combination withother therapeutic agents, for example for use in the treatment of viralinfections. Thus, a compound of this invention can be used incombination with a nucleoside analog such as azidothymidine ordideoxyinosine, atetrahydroimidazo[4,5,1-jk][1,4]-benzodiazepin-2(1H)-one derivative,other HIV reverse transcriptase inhibitors, HIV protease inhibitors, orHIV tat-gene function inhibitors for the prophylaxis against ortreatment of HIV infections. A method for treating a mammal infectedwith a virus comprises administering to a mammal in need of suchtreatment an antivirally effective quantity of a composition comprisingan acceptable pharmaceutical carrier and an antivirally active compoundor compounds or a pharmaceutically acceptable salt thereof.

Furthermore, the non-inflammatory agonists among the compounds of thisinvention may be used to achieve desired physiological results such asinterferon release, interleukin induction, tumor necrosis factorproduction, immune system stimulation and/or reconstitution, insulinsecretion, insulinomimetic activity, acceleration of wound healing,improvement in central nervous system functions such as memory andlearning and abrogation of the symptoms or progress of Alzheimer'sdisease, and any other application for which desirable actions ofprotein kinase C are found.

As phorboid receptor subtype- and/or protein kinase C subtype-selectiveligands, the compounds of this invention also have very valuableapplication as experimental agents for research into the role of proteinkinase C and/or phorboid receptors in important biological processes andin human and veterinary diseases. Thus, their value extends to their useas pharmacological tools for in vitro and in vivo research, in a mannersimilar to the important roles that selective agonists and antagonistshave played in the studies of the mechanism of action of adrenergic,dopaminergic, opiate, benzodiazepine, cholinergic, and serotoninergicreceptor systems, among others.

In addition, the compounds can be used in in vitro diagnostics (e.g., inan assay for protein kinase C). They are also useful as intermediates inthe production of other drugs, e.g., as described in the presentinvention.

The compounds of this invention are generally administered to animals,including but not limited to fish, avians, and mammals including humans.

The pharmacologically active compounds of this invention can beprocessed in accordance with conventional methods of galenic pharmacy toproduce medicinal agents for administration to patients, e.g., mammalsincluding humans.

The compounds of this invention can be employed in admixture withconventional excipients and carriers, i.e., pharmaceutically acceptableorganic or inorganic carrier substances suitable for parenteral, enteral(e.g., oral) or topical application which do not deleteriously reactwith the active compounds. Suitable pharmaceutically acceptable carriersinclude but are not limited to water, salt solutions, alcoholics, gumarabic, vegetable oils, benzyl alcohols, polyethylene glycols, gelatine,carbohydrates such as lactose, amylose or starch, magnesium stearate,talc, silicic acid, viscous paraffin, perfume oil, fatty acid esters,hydroxy methylcellulose, polyvinyl pyrrolidone, etc. The pharmaceuticalpreparations can be sterilized and if desired mixed with auxiliaryagents, e.g., lubricants, preservatives, stabilizers, wetting agents,emulsifiers, salts for influencing osmotic pressure, buffers, coloring,flavoring and/or aromatic substances and the like which do notdeleteriously react with the active compounds. They can also be combinedwhere desired with other active agents, e.g., enzyme inhibitors, toreduce metabolic degradation. Such carriers do not include the followingsolvents when used alone: dimethylsulfoxide, acetone, or methanol orethanol of greater than 80% concentration in water.

For parenteral application, particularly suitable are injectable,sterile solutions, preferably oily or aqueous solutions, as well assuspensions, emulsions, or implants, including suppositories. Ampoulesare convenient unit dosages.

For enteral application, particularly suitable are tablets, dragees,liquids, drops, suppositories, or capsules. A syrup, elixir, or the likecan be used wherein a sweetened vehicle is employed.

A preferred method of administration comprises oral dosing, withtablets, dragees, liquids, drops, or capsules. For the oral route ofadministration, either compounds of this invention lacking functionalgroups destroyed by acid, or tablets or capsules which protect theactive compound from upper gastrointestinal acidity, are preferred.

Sustained or directed release compositions can be formulated, e.g., inliposomes or in compositions wherein the active compound is protectedwith differentially degradable coatings, e.g., by microencapsulation,multiple coatings, absorption onto charcoal, entrapment in human serumalbumin microspheres, etc. It is also possible to freeze-dry the newcompounds and use the lyophilizates obtained, for example, for thepreparation of products for injection.

Another preferred route of administration comprises topical application,for which are employed nonsprayable forms, viscous to semi-solid orsolid forms comprising a carrier compatible with topical application andhaving a dynamic viscosity compatible with topical application,preferably greater than water. Suitable formulations include but are notlimited to solutions, suspensions, emulsions, creams, ointments,powders, liniments, salves, aerosols, etc., which are, if desired,sterilized or mixed with auxiliary agents, e.g., preservatives,stabilizers, wetting agents, buffers or salts for influencing osmoticpressure, etc. For topical application, also suitable are sprayableaerosol preparations wherein the active ingredient, preferably incombination with a solid or liquid inert carrier material, is packagedin a squeeze bottle or in admixture with a pressurized volatile,normally gaseous propellant, e.g., a freon.

The compounds of this invention, admixed with appropriate carriers, mayalso be delivered to subjects by means of an externally connected orinternally implanted pumping device to give a controlled and/orsustained release of the therapeutic mixture, or by means of a patch ofnatural or synthetic fabric and/or polymer impregnated with thecompounds in a suitable carrier and affixed to the skin to achievetransdermal release and absorption of the active compounds.

The compounds of this invention may also be modified by covalentattachment of metabolically modifiable groups, to form "prodrugs" whichare released by cleavage in vivo of the metabolically removable groups.For example, amine, hydroxy and/or thiol groups present in manycompounds of this invention may be converted to prodrugs by covalentattachment of acyl or aminoacyl organic functional groups. Likewise,compounds of this invention containing carboxylic, sulfonic, phosphoric,phosphonic or related free acids, including those in which one or moreoxygen atoms are replaced by sulfur, may be converted to prodrugs byformation of their esters or amides by covalent attachment of alcohols,amines, amino acids and the like. Compounds of this invention may alsoincorporate N-alkyldihydropyridine functional groups, which becomelocalized to the central nervous system after administration to thesubject and subsequent metabolic modification of theN-alkyldihydropyridine group in the central nervous system.

It will be recognized by persons with ordinary skill in medicinalchemistry that conversion of alcohol-, amine-, thiol- or acid-containingcompounds of this invention to prodrugs is preferably done byderivatization of such groups located in regions of the molecule havingminimal steric hindrance, to permit access of metabolizing enzymes,other bioreactants or water. Such alcohol-, amine-, thiol- oracid-containing groups may be located in any of the parent, side-chainor capping-group organic functional groups described above. Aprodrug-type group such as the N-alkyldihydropyridine group, however, ispreferably added to the parent structures P₁ -P₆, etc., unless it isfound for a given compound that the N-alkyldihydropyridine group and/orits metabolic pyridinium oxidation product has desirable bioactivitywhen located in the side chain (S_(o) or S_(x)) or capping group (E_(o)or E_(x)) of a compound of this invention.

It will be appreciated that starting materials for obtaining compoundsof this invention from natural sources or from total or partialsynthesis may be altered in very diverse ways, consistent with thisinvention, to obtain compounds with novel and diverse primarybiological/medicinal activities resulting from, and controlled by, thespecific new S_(o) E_(o) or S_(x) E₁ group created; such propertiesinclude, for example, loss of skin inflammatory activity and appearanceor retention of anti-inflammatory, anti-HIV, anti-leukemic andcytokine-induction activities. It is also possible to introduce anextremely wide variety of changes into either (i) the groups replacingthe hydroxymethyl/1-hydroxyethyl group or (ii) the previously-describedparent phorboid structures, to obtain new entities with improvedsecondary properties, such as, variously, hydrophobicity, watersolubility, potency, oral availability, metabolic and chemicalstability, reduced therapeutic side effects, and so on, using strategiesand techniques widely recognized in the art of medicinal chemistry andpharmacology.

Starting materials for the synthesis of the compounds of this inventionmay be obtained from any of a wide variety of natural sources, and thediterpene, indole alkaloid, diacylglycerol, diaminobenzyl alcohol andpolyacetate compounds are available by total synthesis.

The diversity of phorboids modified in the parent portion and in thehydroxymethyl/1-hydroxyethyl-modified phorboids are illustrated in theExamples, and are also discussed below, separately for each phorboidparent class.

Routine synthetic routes, transformations and procedures common in theart of synthetic organic chemistry are sufficient for highly varied andextensive practice of this invention in great breadth. Indeed, aparticularly remarkable aspect of this invention is the ease with whichhydroxymethyl/1-hydroxyethyl- modified phorboids with highly novelprotein kinase C-modulatory, anti-inflammatory and anti-viralproperties, but lacking skin inflammatory activity, may be obtained fromwell-known starting materials through simple organic chemicaltransformation. This is illustrated by the diversity of Examples foundbelow and by a general discussion, as follows. Although somewhatdifferent kinds of hydroxymethyl/1-hydroxyethyl modifications arediscussed immediately below for the different parent classes ofphorboids, it should be noted that procedures for modification of thehydroxymethyl/1-hydroxyethyl group of one parent class of phorboid aregenerally applicable to the hydroxymethyl/1-hydroxyethyl groups of otherparent classes of phorboids.

Compounds of this invention from the diterpenoid (P₁) class of phorboidsmay be obtained by semisynthetic procedures, starting from any of avariety of compounds from naturally occurring sources as described inthe literature [see Naturally Occurring Phorbol Esters, ed. F. J. Evans,CRC Press, Boca Raton (1986), chapters 7, 8 and 9 and references citedtherein].

Furthermore, these diterpene phorboids are available by total synthesisfrom common organic chemical starting materials. These syntheses providea variety of approaches and associated flexibility in arriving atdiverse functionalities on the parent nucleus and on the final S_(o)E_(o) or S_(x) E_(x) side chain [see Paquette, L. et al., J. Am. Chem.Soc. 106, 1446-1454 (1984); Rigby, J. and Moore, T., J. Org. Chem. 55,2959-2962 (1990); Wender, P. et al., J. Am. Chem. Soc. 111, 8954-8957(1989); Wender, P. et al., J. Am. Chem. Soc. 111, 8957-8958 (1989); andWender, P. and McDonald, F., J. Am. Chem. Soc. 112, 4956-4958 (1990)].

The means for modifying the hydroxymethyl group of diterpenoid phorboidsto produce the compounds of this invention will be obvious to workerswith ordinary skill in synthetic organic chemistry.

In many diterpenoid phorboids the hydroxymethyl group may be modifiedwithout the necessity of protecting other functional groups in themolecule. In other cases, especially when strong nucleophiles or basesare used, other regions of the molecule must be suitably protected usingmethodologies widely practiced in synthetic organic chemistry.Frequently one or more oxygen atoms must be blocked before some types ofchemical modifications may be accomplished on the hydroxymethyl group oron other portions of the diterpene parent. Many widely used andthoroughly characterized protecting groups for the oxygen atoms presentas hydroxy groups are acyl, benzyl, trialkylsilane, benzyloxycarbonyl,4'-methoxyphenyldiphenylmethyl and trimethylsilylyethoxycarbonyl, whichare variously stable to or removed under acidic, basic or reducingconditions or with fluoride ion reagents. The use of such groups isobvious to any worker with modest skill in the art of synthetic organicchemistry. Carbonyl functions may be protected by conversion to acetalsor ketals, or by reduction to the alcohol level followed by protectionwith standard protecting groups for the hydroxy group. With or withoutsuch protection of non-hydroxymethyl portions of the parent diterpenethe hydroxymethyl may, for example, be conveniently capped under verymild conditions by treatment with a substituted or unsubstituted alkyl,aryl, or aralkyl isocyanate, optionally containing silicon or phosphorusatoms in a variety of functional groups, in the presence of a catalystsuch as dibutyltin dilaurate. The resulting compounds lack the toxicinflammatory activity of the phorboid from which they were derived andhave, themselves, anti-inflammatory utility. Illustrations are providedin the Examples below.

Conversion of the hydroxy group to a halogen or pseudohalogen not onlyin itself provides active and useful compounds, but also permits thedisplacement of the resultant electrophile from the protected or in somecases, unprotected, parent nucleus by a very wide range of nucleophiles.Persons with ordinary skill in the art of organic synthesis willrecognize that such nucleophiles may include, without limitation,reagents having carbon, hydrogen, oxygen, nitrogen, sulfur, phosphorus,silicon, arsenic, boron, and/or selenium atoms in their structures.Particular examples would be reaction with ammonia, methylamine, thesodium salt of dimethylphosphine, trimethylphosphite,triphenylphosphine, potassium sulfite, trimethylsilylmethyl-metal salts,lithium trimethylsilylacetylide, sodium cyanide, N-methyl-2-hydroxyethylamine, 1[H]-tetrazole, or with the sodium salt of 2-mercaptoethanol,3-mercaptoethanol, or of hydroxymethylphenol. Many variations may beexecuted as described for nucleophilic displacement of electrophilicgroups in organic molecules by standard textbooks of synthetic organicchemistry, such as J. March, Advanced Organic Chemistry, Third Edition,Wiley-Interscience, New York, 1985. As an illustration,20-deoxy-20-chlorophorbol 12,13-bis-diphenylphosphate yields20-deoxy-20-(pentadecafluoro-[1H,1H]-octylthio)phorbol 12-butyrate13-diphenylphosphate upon treatment withsodiopentadecafluoro-[1H,1H]-octylthiol. As another example, the20-phosphonic acid derivative of 20-deoxyresiniferonol9,13,14-orthophenylacetate may be made by reacting the sodium salt ofdibenzylphosphonate with 20-deoxy-20-bromoresiniferonol9,13,14-orthophenylacetate to form 20-deoxy-20-dibenzylphosphonylresiniferonol 9,13,14-orthophenylacetate dibenzylphosphonyl followed bycatalytic hydrogenolysis of the benzyl ester groups to yield the freephosphonic acid derivative.

By use of protecting groups or modifications of the diterpenoid parentusing methods well-known in the art of synthetic chemistry, the hydroxygroup of the hydroxymethyl group may be replaced by a metal and thenreacted with an electrophile, effectively replacing the hydroxy with agroup derived from the electrophile. The techniques for this replacementare obvious to workers with ordinary skill in organic synthesis, in thatreplacement of the hydroxymethyl hydroxy group by halogen in a suitablyprotected or modified diterpene permits strong and/or hard nucleophilesto be generated by the use of metals or strong bases, and persons withordinary skill in the art of organic chemistry will recognize that suchnucleophiles can be contacted with a very diverse range of electrophilicreagents having carbon, hydrogen, oxygen, nitrogen, sulfur, phosphorus,silicon, arsenic, boron and/or selenium atoms in their structures toobtain hydroxymethyl-modified diterpenes of widely varying structures.As one approach among many, the hydroxymethyl group of an appropriatelyprotected diterpene may be converted to the 20-bromo derivative withmethanesulfonyl bromide in dimethylformamide/pyridine. The resultingbromide may be modified by halogen-metal exchange using appropriatelyactive metals or metal-containing reagents such as magnesium, zinc,alkali metals, metal alkyl reagents and so on, to obtain carbanioniccharacter at the carbon atom previously bearing the hydroxy group, asshown in the following structures, which also illustrate typicalprotection of oxygen functions elsewhere in the molecule: ##STR38##wherein Met is the metal ion, PG₁ -PG₃ are typically selected from thelist above and PG₄ is typically 2',2'-propylidene or dialkyl- ordiarylsilyl. PG₃ may be unnecessary in some cases (i.e. may be hydrogenor an anionic charge). The simultaneous presence or subsequent additionto the metal/anion-containing reaction of an electrophile such as,without limitation, an aldehyde, ketone, epoxide or oxetane thenprovides, after reaction and workup, compounds having one or moremethylenes inserted between the original hydroxy group and the methyleneto which it was attached. This methodology is particularly advantageousfor replacement of the hydroxy with a group containing silicon,phosphorous or other atoms by contacting the anionic metal derivative ofthe parent nucleus with electrophilic reagents having halogen orpseudohalogen groups, in addition to other functions chosen forbiological specificity, bonded directly to the silicon or phosphorusatoms, affording compounds with useful biological activity.

For those diterpene-type phorboids wherein the hydroxy of thehydroxymethyl group is allylic, such as phorbol, ingenol, andresiniferonol esters, the replacement of the hydroxy by chloro orpreferably by bromo or iodo yields compounds that can be convenientlyreacted with activated zinc in the presence of electrophiles such as,without limitation, aldehydes, ketones, epoxides, or oxetane; theresultant compounds have one or more methylenes inserted between theoriginal hydroxy group and the methylene to which it was attached, or,after an allylic rearrangement, a new functional group results atposition 7 of the diterpene skeleton. An illustration of this would bethe reaction of 20-deoxy-20-chlorophorbol 12-myristate 13-acetate withan excess of formaldehyde and an excess of zinc in the presence oftetrahydrofuran and saturated ammonium chloride solution with vigorousstirring for 24 hours. The resultant compound has the 6,7-double bondrearranged to the 6,20-position and bears a new hydroxymethyl groupattached to position 7 instead of to position 6 as in the parentditerpene, and this compound,12-beta-myristoyloxy-13-acetoxy-4,9-dihydroxy-7-hydroxymethyl-1,6(20)-tigliadien-3-one,has good anti-inflammatory activity.

Alternatively, the hydroxy group on the hydroxymethyl of appropriatelyprotected or modified diterpenoids may be oxidized to an aldehyde andthen reacted via condensation or addition chemistries to provide a verywide variety of modified indolactams. Examples, without limitation,would be reactions with Wittig reagents; hydroxylamines; hydrazines;semicarbazides; ammonia, primary amines or secondary amines followed bysodium cyanoborohydride reduction to primary amines, secondary amines ortertiary amines, respectively; or Grignard reagents. Many variations maybe executed as described in standard textbooks of synthetic organicchemistry, such as J. March, op cit. After completion of reactions inthe hydroxymethyl region, an acetonide group protecting the 3-OH and4-OH groups is removed with mild acid followed by reoxidation of the3-OH to keto with sulfur trioxide-pyridine-dimethylsulfoxide.Trialkylsilane protecting groups PG₁ and PG₂ may then be removed withfluoride ion, preferably followed by derivatization of the now-free 12-and 13-OH's with acylating agents, isocyanates, alkyl- or arylchloroformates or alkylating agents to establish hydrophobic groups inthis region of the diterpene.

Even a relatively base-sensitive compound such as a phorbol12,13-diester may be oxidized to the 20-aldehyde with manganese dioxideand reacted directly, as is obvious to workers with ordinary skill insynthetic organic chemistry, with stabilized Wittig reagents such asmethyl triphenylphosphoranylideneacetate, to yield a20-deoxy-20-methoxycarbonylmethylidenephorbol 12,13-diester.

Alternatively, a phorbol 12,13-diester may be selectively protected atthe 4 and 9 hydroxy groups using trimethylsilyltrifluoromethanesulfonate, followed by reaction with manganese dioxideto obtain a protected aldehyde. The latter compound may be successfullytreated with strong Wittig reagents such as the lithium salt of2-hydroxyethylidenetriphenylphosphorane, followed by deprotection withtetrabutylammonium fluoride to obtain the corresponding chain-extended,20,21-didehydro-21,22-dihomophorbol diester.

The hydroxymethyl group of suitable diterpene phorboids may be oxidizedto the carboxylic acid level by methods well-known in the art or thesecarboxylic acids may be prepared by modifications of the total synthesesdescribed in the literature cited above. Besides being useful examplesof this invention themselves these acids permit the preparation offurther modified diterpene phorboids. For example, this carboxylic groupmay be activated for condensation reactions by any of a number ofwell-known methods, e.g. by conversion to an acyl halide or to an activeester such as the N-succinimidyl ester. The resultant activated carboxylmay then be easily convened to simple or multifunctional ester, amide,or thioester derivatives by reaction with alcohols, amines, or thiolsrespectively, alone or in the presence of condensation catalysts.

The use of the methods of total synthesis as described in the literaturecited above permits specific modifications of the parent structures ofthe diterpenoids. By established techniques in the art of organicsynthesis modified parent structures may be obtained which embodyalterations at the hydroxymethyl group as well, and which have usefulbiological activity. This wide variety of modified diterpenoidstructures may result from the use of modified starting materials, frommodifications of one or more synthetic steps or from a combination ofboth. Starting materials for the synthesis of the compounds of thisinvention from the indolactam (P₂) class of phorboids may be obtainedfrom any of a variety of natural sources as described in the literature[T. Sugimura, Gann 73 499-507 (1982), H. Fujiki and T. Sugimura,Advances in Cancer Research 49, 223-264 (1987), K. Irie and K.Koshimizu, Mem. Coll. Agric., Kyoto Univ. 132, 1-59 (1988) andreferences cited therein]. Furthermore, these indolactam phorboids areavailable by total synthesis from common organic chemical startingmaterials. These syntheses provide a considerable variety of approachesand associated flexibility in arriving at highly diverse functionalitieson the parent nucleus and on the final S_(o) E_(o) or S_(x) E_(x) sidechain, and include, but are not limited to, the procedures described byY. Endo et al., Tetrahedron 42, 5905-5924 (1986), S. de Laszlo et al.,J. Chem. Soc., Chem. Comm. 344-346 (1986), S. Nakatsuka et al.,Tetrahedron Letters 27, 5735-5738 (1986) and Tetrahedron Letters 28,3671-3674 (1987), H. Muratake and M. Natsume, Tetrahedron Letters 28,2265-2268 (1987), M. Mascal and C. Moody, J. Chem. Soc., Chem. Comm.589-590 (1988), A. Kozikowski et al., J. Am. Chem. Soc. 111, 6228-6234(1989), and T. Kogan et al., Tetrahedron 46, 6623-6632 (1990) andreferences therein.

Given indolactams containing hydroxymethyl groups, the means formodifying the hydroxymethyl group to produce the compounds of thisinvention will be obvious to workers with ordinary skill in syntheticorganic chemistry.

In some cases, the hydroxymethyl group may be modified once otherregions of the molecule have been suitably protected using simple,obvious and widely-precedented methodologies practiced extensively insynthetic organic chemistry. In some specific cases the indole nitrogenmust be blocked before some types of chemical modifications may beaccomplished on the hydroxymethyl group or on other portions of theindolactam parent. Many protecting groups for the indole nitrogen havebeen used in the organic chemical literature, and their use here isobvious. Among these are t-butoxycarbonyl, acetyl, benzyl,trimethylsilylyethoxymethylene, benzyloxycarbonyl and benzenesulfonyl,which are variously stable to or removed under acidic, basic or reducingconditions or with fluoride ion reagents. With or without suchprotection the hydroxymethyl may, for example, be conveniently cappedunder very mild conditions by the treatment with a substituted orunsubstituted alkyl, aryl, or aralkyl isocyanate, optionally containingsilicon or phosphorus atoms in a variety of functional groups, in thepresence of a catalyst such as dibutyltin dilaurate. The resultingcompounds lack the toxic inflammatory activity of the phorboid fromwhich they were derived, and have themselves anti-inflammatory utility.As an illustration, treatment of (-)-7-octylindolactam V withmethylisocyanate in the presence of dibutyltin dilaurate and4-dimethylaminopyridine in tetrahydrofuran affords14-O-(N-methyl)carbamoyl-7-octyl-(9S,12S)-indolactam V.

Conversion of the hydroxy group to a halogen or pseudohalogen not onlyin itself provides active and useful compounds, but also permits thedisplacement of the resultant electrophile from the protected or in somecases, unprotected, parent nucleus by a very wide range of nucleophiles.Persons with ordinary skill in the art of organic synthesis willrecognize that such nucleophiles may include, without limitation,reagents having carbon, hydrogen, oxygen, nitrogen, sulfur, phosphorus,silicon, arsenic, boron, and/or selenium atoms in their structures.Particular examples would be reaction with ammonia, methylamine, thesodium salt of dimethylphosphine, trimethylphosphite,triphenylphosphine, potassium sulfite, trimethylsilylmethyl-metal salts,lithium trimethylsilylacetylide, sodium cyanide, N-methyl-2-hydroxyethylamine, 1[H]-tetrazole, or with the sodium salt of 2-mercaptoethanol,3-mercaptoethanol, or of hydroxymethylphenol. Many variations may beexecuted as described in standard textbooks of synthetic organicchemistry, such as J. March, Advanced Organic Chemistry, Third Edition,Wiley-Interscience, New York, 1985. As an illustration,14-O-methanesulfonyl-1-trimethylsilylmethylindolactam V yields14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-1-trimethylsilylmethylindolactamV upon treatment with sodiopentadecafluoro-[1H,1H]-octylthiol. In asimilar manner14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-6,7-tetramethyleneindolactamV is prepared. See Y. Endo et al., Tetrahedron 42, 5905-5924 (1986) fora synthesis of 6,7-tetramethyleneindolactam V. As a furtherillustration, treatment of14-O-methanesulfonyl-7-[2'-(2"-diphenylphosphonylethyl)aminoethyl]indolactamV with the sodium salt of 4-t-butyldimethylsilyloxy-2-phenylphenolfollowed by treatment of the product with tetrabutylammonium fluoride intetrahydrofuran affords14-O-(4'-hydroxy-2'-phenyl)phenyl-7-[2'-(2"-diphenylphosphonylethyl)aminoethyl]indolactamV. In a similar manner14-O-(5'-hydroxy-1'-naphthyl)-7-[1',1'-dimethyl-3'-(hexyldimethylsilyl)propyl]-indolactamV, 14-O-(3'-hydroxy-5'-benzyloxy)phenyl-7-octylindolactam V,14-O-(4'-hydroxy-2'-phenyl)phenyl-6,7-tetramethyleneindolactam V,14-O-(5'-hydroxy-1'-naphthyl)-6,7-tetramethyleneindolactam V,14-O-(3'-hydroxy-5'-benzyloxy)phenyl-6,7-tetramethyleneindolactam V areprepared. As a further illustration, treatment of14-O-methanesulfonyl-7ooctylindolactam V with sodium sulfite in methanolprovides 14-deoxy-7-octylindolactam V 14-sulfonic acid. In a similarmanner 14-deoxy-6,7-tetramethyleneindolactam V 14-sulfonic acid isprepared. As another example, the 14-phosphonic acid derivative of14-deoxy-6,7-tetramethyleneindolactam V may be made by reacting thesodium salt of dibenzylphosphonate with14-O-methanesulfonyl-7-octylindolactam V to form14-deoxy-14-dibenzylphosphonyl-6,7-tetramethyleneindolactam V followedby catalytic hydrogenolysis of the benzyl ester groups to yield the freephosphonic acid.

By use of protecting groups or modifications of the indolactam parent bymethods well-known in the art of synthetic chemistry, the hydroxy groupof the hydroxymethyl may be replaced by a metal and then reacted with anelectrophile, effectively replacing the hydroxy with a group derivedfrom the electrophile. The techniques for this replacement are obviousto workers with ordinary skill in organic synthesis, in that replacementof the hydroxymethyl hydroxy group by halogen in a suitably protected ormodified indolactam permits strong and/or hard nucleophiles to begenerated by the use of metals or strong bases, and persons withordinary skill in the art of organic chemistry will recognize that suchnucleophiles can be contacted with a very diverse range of electrophilicreagents having carbon, hydrogen, oxygen, nitrogen, sulfur, phosphorus,silicon, arsenic, boron and/or selenium atoms in their structures toobtain hydroxymethyl-modified indolactams of widely varying structures.As one approach among many, the 14-O-methanesulfonyl derivative of anappropriate indolactam may be converted to an iodo compound with sodiumiodide in acetone. The resulting iodide may be modified by halogen-metalexchange using appropriately active metals or metal-containing reagentssuch as magnesium, zinc, alkali metals, metal alkyl reagents and so on,to obtain carbanionic character at the carbon atom previously bearingthe hydroxy group, as shown: ##STR39## wherein Met is the metal ion, PG₅may be one of the indole nitrogen protecting groups listed above, andPG₆ may be unnecessary (i.e. may be hydrogen or an anionic charge), maybe benzyl (removable by hydrogenolysis) or may be a stable group, suchas methyl or ethyl, intended to remain in the final bioactive syntheticproduct. The simultaneous presence or subsequent addition to thereaction of an electrophile such as, without limitation, an aldehyde,ketone, epoxide or oxetane then provides, after reaction and workup,compounds having one or more methylenes inserted between the originalhydroxy group and the methylene to which it was attached. Thismethodology is particularly advantageous for replacement of the hydroxywith a group containing silicon, phosphorous or other atoms bycontacting the anionic metal derivative of the parent nucleus withelectrophilic reagents having halogen or pseudohalogen groups, inaddition to other functions chosen for biological specificity, bondeddirectly to the silicon or phosphorus atoms, affording compounds withuseful biological activity.

Alternatively, the hydroxy group on the hydroxymethyl of appropriatelyprotected or modified indolactams may be oxidized to an aldehyde andthen reacted via condensation or addition chemistries to provide a verywide variety of modified indolactams. Examples, without limitation,would be reactions with Wittig reagents, hydroxylamines, or Grignardreagents. Many variations may be executed as described in standardtextbooks of synthetic organic chemistry, such as J. March, op cit. Asan illustration, the aldehyde, 14-deoxy-14-oxo-7-octylindolactam V, maybe prepared by oxidation of the parent hydroxymethyl compound withperiodinane among other oxidizing agents. This aldehyde may be treatedwith O-(4-mercaptophenyl)hydroxylamine hydrochloride in ethanol toafford the O-(4-mercaptophenyl)oxime. This aldehyde may also be treatedwith N¹ -(3-methoxyphenyl)semicarbazide to afford the corresponding N¹-(3-methoxyphenyl)semicarbazone. The O-(4-mercaptophenyl)hydroxylaminehydrochloride may be prepared from O-(4-nitrophenyl)hydroxylaminehydrochloride [A. Hirsch et al., J. Med. Chem. 20, 1546-1551 (1977)] inways obvious to one with ordinary skill in the art of organic synthesis.

These aldehydes may also be obtained by reduction of appropriatecarboxylic acid derivatives by application of well-known techniques. Asan illustration,9-deshydroxymethyl-9-methoxycarbonyl-6,7-tetramethyleneindolactam V(prepared as described below) may be reduced to the aldehyde,14-deoxy-14-oxo-6,7-tetramethyleneindolactam V, by any of severalreagents widely known in the art of organic chemistry, for examplediisobutylaluminum hydride. This aldehyde may be further modified toafford the O-(4-mercaptophenyl)oxime and the N¹-(3-methoxyphenyl)semicarbazone in the manner described above.

The hydroxymethyl group of suitable indolactam phorboids may be oxidizedto the carboxylic acid level by methods well-known in the art or thesecarboxylic acids may be prepared by modifications of the total synthesesdescribed in the literature cited above. Besides being useful examplesof this invention themselves these acids permit the preparation offurther modified indolactams. For example, this carboxylic group may beactivated for condensation reactions by any of a number of ordinary andwell-known methods, e.g. by conversion to an acyl halide or to an activeester such as the N-succinimidyl ester. The resultant activated carboxylmay then be easily converted to simple or multifunctional ester, amide,or thioester derivatives by reaction with alcohols, amines, or thiolsrespectively, alone or in the presence of condensation catalysts. As anillustration, rac-9-deshydroxymethyl-9-carboxyindolactam V (obtained asdescribed below) may be treated with N-hydroxysuccinimide anddicyclohexylcarbodiimide in acetonitrile. After purification, theresulting N-succinimidyl ester may be treated with3-amino-1,2-propandiol and 4-dimethylaminopyridine in methylenechloride/tetrahydrofuran to affordrac-9-deshydroxymethyl-9-[N-(2',3'-dihydroxy)propyl]carboxamidoindolactamV. The stereoisomers may be obtained separately either by usingoptically active starting materials or by separation from the mixture bychromatography on an enantioselective column packing [D. Armstrong,Analytical Chem. 59. 84-91A (1987)].

The total syntheses described in the literature cited above are alsoamenable to extensive adaptations so as to provide a wide variety ofmodifications in the parent structures of the indolactam group. Byestablished techniques in the art of organic synthesis modified parentstructures may be obtained which embody alterations at the hydroxymethylgroup and which have useful biological activity. This extremely widevariety of modified indolactam structures may result from the use ofmodified starting materials, from modifications of one or more syntheticsteps or from a combination of both. As an illustration,9-deshydroxymethyl-9-carboxy-6,7-tetramethyleneindolactam V may beprepared from 4-nitro-6,7-tetramethylenegramine (Y. Endo et al., opcit.) by the application of several routes obvious to workers withordinary skill in organic synthesis. For example, methylα-nitro4-amino-6,7-tetramethyleneindole-3 acetate is prepared from4-nitro-6,7-tetramethylenegramine in the manner described by T. Masudaet al., Agric. Biol. Chem. 53, 2257-2260 (1989). Alkylation of the aminogroup of this compound with benzylD-α-trifluoromethylsulfonyloxyisovalerate in the manner described by T.Kogan etal., op cit., affords an optically active intermediate which,after removal of the benzyl ester and reduction of the nitro group, islactamized in the manner of T. Masuda et al., op cit., and methylated bytreatment with formaldehyde/sodium cyanoborohydride to afford9-deshydroxymethyl-9-methoxycarbonyl-6,7-tetramethylene-(9S,12S)-indolactamV. Hydrolysis with methanolic sodium hydroxide affords9-deshydroxymethyl-9-carboxy-6,7-tetramethylene-(9S,12S)-indolactam V.

To further illustrate the synthesis of the compounds of this invention,the modified indolactam,1,2,4,5,6,8-hexahydro-5-methyl-2-(1-methylethyl)-3H-pyrrolo(4,3,2-gh)-1,4-benzodiazonin-3-one,may be prepared from N-BOC-4-nitrotryptophanol [Y. Endo et al.,Tetrahedron 42, 5905-5924 (1986)] by the application of several routesobvious to workers with ordinary skill in synthetic chemistry. Forexample, preparation of N-BOC-4nitrodeoxytryptophanol may beaccomplished by reduction of the phenylselenium derivative withtriphenyltin hydride [D. Clive et al. Chemical Communications 41-42(1978)] or by reduction of the mesylate derivative with lithiumtriethylborohydride [R. W. Holder and M. G. Matturro, J. Org. Chem. 42,2166-2168 (1977)] or by several other routes. From the deoxy derivativethe synthesis could proceed in the manner described by Y. Endo et al.,loc cit., for the hydroxy derivative. Specifically, the resultingsubstituted indolylvaline methyl ester is hydrolyzed and the resultingacid is converted to the N-succinimidyl ester. Upon cleavage of the BOCgroup under acidic conditions cyclization to the lactam occurs directlyto provide all four stereoisomers, i.e., 2R,5R-, 2R,5S-, 2S,5S-, and2S,5R-1,2,4,5,6,8-hexahydro-5-methyl-2-(1-methylethyl)-3H-pyrrolo(4,3,2-gh)-1,4-benzodiazonin-3-one. These stereoisomers may be obtainedseparately either by beginning the synthesis with optically activeN-BOC-4-nitrotrytophanol or by separation from the mixture bychromatography on an enantioselective column packing [D. Armstrong,Analytical Chem. 59, 84-91A (1987)]. Similarly, the further modifiedindolactam,1-(1-oxobutyl)-1,2,4,5,6,8-hexahydro-5-methyl-3H-pyrrolo(4,3,2-gh)-1,4-benzodiazonin-3-one,may be prepared. Specifically, hydrogenation ofN-BOC-4-nitrodeoxytrytophanol over palladium on carbon will provideN(2')-BOC-4-aminodeoxytrytophanol. Alkylation of this compound withmethyl bromoacetate affords N-[3-(N-BOC-2'-aminopropyl)4-indolyl]glycinemethyl ester [Y. Endo, et al., Chem. Pharm. Bull. 30, 3457-3460 (1982)].Acylation of this material with butanoyl chloride in the presence ofpotassium carbonate or pyridine will provideN-butanoyl-N-[3-(N-BOC-2-aminopropyl)-4-indolyl]glycine methyl ester.This latter material may be converted to1-(1-oxobutyl)-1,2,4,5,6,8-hexahydro-5-methyl-3H-pyrrolo(4,3,2-gh)-1,4-benzodiazonin-3-oneby application of the methods of Y. Endo, et al., loc cit. (1986). Theenantiomers, 5R- and 5S-, may be obtained separately by beginning withoptically active materials as described above or by separation at thefinal stage by chromatography also as described above.

Derivatives of the indole, indene, benzofuran and benzothiophene classesin which the benzenoid ring contains one or two nitrogen atoms inpositions 5, 6 and/or 7 may be obtained by modifying the known syntheticsequences for indolactams cited above. For example, a 5-, 6- and/or7-monoaza- or diaza-indolactam derivatives may be obtained by replacingthe 4-aminoindole starting material in an appropriate synthetic sequencewith a 5-, 6- and/or 7omonoaza- or diazaindole.

Beyond the extensive changes in the hydroxymethyl group of indolactamand related P₂ -type parent phorboids described above, the presentinvention also discloses broad and diverse alterations which areaccommodated in the non-hydroxymethyl regions of the parent P₂ -typephorboids. Such modifications of the indolactam parents can be carriedout before, after or in alternating fashion with respect to constructionof the hydroxymethyl modifications, depending on obvious considerationsof chemical stability of the various functional groups in intermediatesbeing subjected to chemical modifications.

It is obvious to one skilled in the art that many modified indolactamparents may be obtained by carrying a modification through the de novosynthesis as described in the literature cited above. As anillustration, 7-alkylindolactam Vs containing a wide variety of alkylgroups may be prepared by application of the method of A. Kozikowski etal., op cit., to a variety of alkyl substituted isoxazolines. As afurther illustration, the modified indolactam,1-(1'-octyl)-1,2,4,5,6,8-hexahydro-5-hydroxymethyl-3H-pyrrolo[4,3,2-gh]-1,4-benzodiazonin-3-one(13-N-octylindolactam G), may be prepared. Alkylation of 4-aminoindolewith methyl bromoacetate followed by acylation with octanoyl chloride inthe presence of pyridine affords N-octanoyl-N-(4-indolyl)glycine methylester. This is converted to 13-N-octylindolactam G by the methoddescribed by A. Kozikowski et al., op cit. The enantiomers are obtainedseparately by silica gel chromatography of the14-O-[N-(S)-(1'-naphthyl)ethyl]carbamates followed by reduction withtrichlorosilane-triethylamine. Further modifications of this material toproduce hydroxymethyl modified embodiments of this invention are carriedout by the preparation of 1-N-(t-butyloxycarbonyl)-13-N-octylindolactamG, according to the method of Y. Endo et al., Tetrahedron 43, 2241-2247(1987). Then,14-O-methanesulfonyl-1-N-(t-butyloxycarbonyl)-13-N-octylindolactam G maybe prepared by treatment of the above indolactam with methanesulfonylchloride. Treatment of this material with a variety of nucleophiles asdescribed above followed by removal of the t-butyloxycarbonyl group bytreatment with acid, for example trifluoroacetic acid in methylenechloride, affords a very wide variety of hydroxymethyl-modifiedderivatives. 14-Deoxy-14-(2'-hydroxyethylthio)-13-N-octylindolactam Gand 14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-13-N-octylindolactamG are specific illustrations of the types of useful compounds which maybe prepared by application of these approaches.

As a further illustration, 1,2-tetramethyleneindolactam V may beprepared. One of the many preparations of this parent uses2-carbethoxy-4-nitroindole, which is a side-product from the preparationof 4-nitroindole, as the starting material for transformation to4-amino-1,2-tetramethyleneindole. Alkylation of the amino group of thiscompound with benzyl D-α-trifluoromethylsulfonyloxyisovalerate in themanner described by T. Kogan et al., op cit., affordsN-(1,2-tetramethyleneindol-4-yl)valine benzyl ester as a singleenantiomer. This material may then be convened to1,2-tetramethyleneindolactam V by the methods described by A. Kozikowskiet al., op cit. Further modifications of this material to producehydroxymethyl modified embodiments of this invention may be carried outby the techniques discussed above. As specific examples,14-deoxy-14-(2'-hydroxyethylthio)-1,2-tetramethyleneindolactam V and14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-1,2-tetramethyleneindolactamV are prepared from 14-O-methanesulfonyl-1,2-tetramethyleneindolactam V.

Further synthetic elaborations may be effected at the end or near theend of the preparation of the modified indolactam-parent or of thehydroxymethyl-modified indolactam. As one of many possibleillustrations, 7-(2'-methylbut-3'-en-2'-yl)indolactam V (pendolmycin),which may be prepared by the method of Kozikowski et al., op cit., or bythe method of Okabe et al., Tetrahedron 46, 5113-5120 (1990), or aderivative of pendolmycin, may be hydroborated. The resultingtrialkylborane, or haloborane, may be converted into other functionalgroups including but not limited to hydroxy, carbonyl, alkyl, amino andhalo, all by well-known methods such as those referenced by A. Pelter etal., Borane Reagents, Academic Press, London, 1988. In a specificillustration, t-butyldimethylsilyl ether of pendolmycin may be preparedby treatment of a dimethylformamide solution of pendolmycin andimidazole (1:10 molar ratio) with t-butyldimethylchlorosilane. Treatmentof this ether with 9-borabicyclo[3.3.1]nonane (9-BBN) affords thetrialkylborane at the terminal carbon of the former terminal olefin, theoxidation of which with hydrogen peroxide in sodium hydroxide affordsthe primary alcohol. Acetylation of this alcohol with acetic anhydridein pyridine and removal of the silyl ether with tetrabutylammoniumfluoride in tetrahydrofuran provides7-(2'-methyl-4'-acetoxybutan-2'-yl)indolactam V. This may then beconverted to14-deoxy-14-(2"-hydroxyethylthio)-7-(2'-methyl-4'-hydroxybutan-2'-yl)indolactamV or many other hydroxymethyl modified derivatives by the methodsdiscussed above followed by treatment with sodium hydroxide in methanolto hydrolyze the acetate, yielding a hydroxy group amenable to a widerange of further transformations, preferably to establish a hydrophobicsubstituent in that position of the molecule.

Similarly, compounds containing silicon and phosphorus may be obtainedby adding reagents having Si--H or P--H bonds across the double bond ofpendolmycin, for example, under the influence of chloroplatinic acid orlight/peroxide catalysis, respectively. Such means for introducingsilicon or phosphorus-containing groups may also be applied tostructures having vinyl groups elsewhere in the molecule. For example,diethylphosphine may be added across the double bond of(-)-9-deshydroxymethyl-9-vinyl-7-octylindolactam V in a reactionpromoted by light and peroxide catalysis to obtain14-deoxy-14-diethylphosphinylmethyl-7-octylindolactam V.

An illustration of the modification of the parent group after theintroduction of appropriate hydroxymethyl modifications is thepreparation of14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-7-[2'-methyl-4'-(N-octanoyl-N-ethyl)aminobutan-2'-yl]indolactamV. Hydroboration of14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)pendolmycin, prepared inthe manner discussed above from 14-methanesulfonylpendolmycin, withdichloroboranedimethylsulfide in the presence of boron trifluorideprovides14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-7-(2'-methyl-4'-dichloroboranylbutan-2'-yl)indolactamV which upon treatment with ethylazide affords14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-7-(2'-methyl-4'-ethylaminobutan-2'-yl)indolactamV, itself amenable to further reactions, for example acylation withoctanoyl chloride to yield the more hydrophobic14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-7-[2'-methyl-4'-N-octanoyl-N-ethyl)aminobutan-2'-yl]indolactamV.

The application of established techniques in the art of syntheticchemistry to naturally derived or to synthetically derived parents ofthe indolactam group also permits the obtainment of specificallymodified parents of that class. These modified parents may then befurther modified at the hydroxymethyl group by the methods discussedabove. To illustrate this, 14-O-t-butlydimethylsilylindolactam V [Y.Endo et al., Tetrahedron 43, 2241-2247 (1987)] may be treated withsodium hydride in dimethylformamide followed by phosgene. The reactivechloroformamide so formed is quenched with perfluoroheptylmethanol toafford, after removal of the silyl ether,1-N-(pentadecafluoro-[1H,1H]-octyloxy)carbonylindolactam V. Byapplication of methods discussed above a variety of hydroxymethylmodified derivatives may be prepared as specifically illustrated by14-deoxy-14-(2'-hydroxyethylthio)-1-N-(pentadecafluoro-[1H,1H]-octyloxy)carbonylindolactamV and by14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-1-N-(pentadecafluoro-[1H,1H]-octyloxy)carbonylindolactamV. As a further illustration, 14-O-acetylindolactam V [K. Irie and K.Koshimizu, Mem. Coll. Agric., Kyoto Univ. 132, 1-59 (1988)] may bealkylated with ethyl acrylate in the presence of sodium hydride toafford 14-O-acetoxy-I-N-(2'-ethoxycarbonyl)indolactam V. Treatment ofthis material with aluminum chloride in nitrobenzene followed byreduction of the purified product with lithium aluminum hydride-aluminumchloride in tetrahydrofuran provides 1,7-trimethyleneindolactam V. Byapplication of methods discussed above a variety of hydroxymethylmodified derivatives may be prepared as specifically illustrated by14-deoxy-14-(2'-hydroxyethylthio)-1,7-trimethyleneindolactam V and by14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-1,7-trimethyleneindolactamV.

Such specifically modified parents of the indolactam class may also befurther modified at positions other than the hydroxymethyl group eitherbefore or after the modifications of hydroxymethyl group to produceembodiments of this invention. The means for accomplishing thesemodifications are obvious to someone with ordinary skill in organicsynthesis. As an illustration, 7-(2'-aminoethyl)indolactam V may beprepared from indolactam V by methods similar to those described by K.Irie and K. Koshimizu, op cit. N-Derivatization of this compound withpentadecafluoro-[1H,1H]-octyl isocyanate, 3-trimethylsilylpropanoylchloride or 3-diphenyloxophosphinylpropanoyl chloride yields7-[2'-(N'-pentadecafluoro-[1H,1H]-octylureido)ethyl]indolactam V,7-[2'-(N-(3"-trimethylsilylpropanoyl)amino)ethyl]indolactam V or7-[2'-(N-(3"-diphenyloxophosphinylpropanoyl)amino)ethyl]indolactam Vrespectively. By application of methods discussed above a variety ofhydroxymethyl modified derivatives may be prepared as specificallyillustrated by14-deoxy-14-azido-7-[2'-(N'-pentadecafluoro-[1H,1H]-octylureido)ethyl]indolactamV,14-deoxy-14-(2'-hydroxyethoxy)-7-[2'-(N-(3"-trimethylsilylpropanoyl)amino)ethyl]indolactamV,14-O-[1'(5'-hydroxynaphthyl)]-7-[2'-(N-(3"-diphenyloxophosphinylpropanoyl)amino)ethyl]indolactamV,14-deoxy-14-mercapto-7-[2'-(N'-pentadecafluoro-[1H,1H]-octylureido)ethyl]indolactamV,14-deoxy-14-(pentadecafluoro-[1H,1H]-octylthio)-7-[2'-(N-(3"-trimethylsilylpropanoyl)amino)ethyl]indolactamV, and14-deoxy-7-[2'-(N-(3"-diphenyloxophosphinylpropanoyl)amino)ethyl]indolactamV 14-sulfonic acid.

As a further illustration, 14-O-acetyl-2-formyl-7-ethylindolactam V maybe prepared by the treatment of 14-O-acetyl-7-ethylindolactam V withdichloromethyl methyl ether-titanium tetrachloride [K. Irie et al., Int.J. Cancer 43 513-519 (1989)]. Condensation with nitromethane followed byreduction of the nitrovinyl derivative with lithium aluminumhydride-aluminum chloride affords 2-(2'-aminoethyl)-7-ethylindolactam V.Derivatization of this compound with pentadecafluoro-[1H,1H]-octylisocyanate, trimethylsilylpropanoyl chloride or3-diphenyloxophosphinylpropanoyl chloride yields7-ethyl-2-[2'-(N'-pentadecafluoro-[1H,1H]-octylureido)ethyl]indolactamV, 7-ethyl-2-[2'-(N-(3"-trimethylsilylpropanoyl)amino)ethyl]indolactam Vor7-ethyl-2-[2'-(N-(3"-diphenyloxophosphinylpropanoyl)amino)ethyl]indolactamV respectively. By application of methods discussed above a variety ofhydroxymethyl modified derivatives may be prepared.

Many parent phorboids of the diaminobenzyl alcohol (P₃) class areavailable via total synthesis [see, for example, Wender, P. et al.,Proc. Nat. Acad. Sci. USA 83, 4214-4218, (1986)]. Given the readyavailability of a wide variety of precursors for phorboids of this classand for hydroxymethyl-modified members of that class, the means forproducing the P₃ -based compounds of this invention will be obvious toworkers with ordinary skill in synthetic organic chemistry.

For example, 4-carboxy-6-(N-decanoylamino)indole may be converted to itsN-hydroxysuccinimide ester by reaction with one equivalent ofdicyclohexylcarbodiimide and one equivalent of N-hydroxysuccinimide inacetonitrile/tetrahydrofuran/methylene chloride suspension. The productN-hydroxysuccinimide ester is purified and then reacted with3-amino-1,2-propanediol in tetrahydrofuran to obtain4-(N-2',3'-dihydroxypropylcarboxamido)-6-(N-decanoylamino)indole.

Given the ready availability of a wide variety of precursors forphorboids of the diacylglycerol (P₄) class and forhydroxymethyl-modified members of that class, the means for producingcompounds of this invention based on the P₄ parent phorboid class willbe obvious to workers with ordinary skill in synthetic organicchemistry. In some cases these precursors will be available withsuitable blocking groups in place or they may be put in place using wellunderstood techniques or blocking groups are not needed. An illustrationof the preparation of compounds of this invention is the preparation of3R-3-carboxy-1,6-dioxo-2,5-dioxacyclotetracosane. Diesterification ofreadily available D-glyceric acid with 9-decynoyl chloride in thepresence of pyridine affords 2,3-O,O-didecynoyl-D-glyceric acid.Treatment of this material with cupric acetate/pyridine affords thematerial coupled at the acetylenic termini, the catalytic hydrogenationof which affords 3R-3-carboxy-1,6-dioxo-2,5-dioxacyclotetracosane. Insimilar and related manner 2R-2,3-octanoyloxypropionic acid and4R-4-carboxy-2,7-dioxo-3,6-dioxa-1,8-diazacyclocosane may be prepared.Furthermore, the same acylation with terminal acetylenic carboxylicacids followed by cyclization and reduction may be applied to1-chloro-2,3-dihydroxypropane, preceded or followed by displacement ofthe chloro leaving group with any of the nucleophilic type of reagentsdescribed above, to yield, after an appropriate sequence ofprotection-deprotection of chemically sensitive functional groups in theintermediates, novel and useful hydroxymethyl-modified phorboids of thediacylglycerol class.

Compounds of the polyacetate (P₅) class of phorboids may be obtained bysemisynthetic modification of starting compounds purified from naturalsources [see Mynderse, J. et al., Science 196, 538-540 (1977) andreferences cited therein] or by adaptations and modifications of theintermediates or final products of total syntheses of the polyacetates[see Park, P. et al., J. Am. Chem. Soc. 109, 6205-6207 (1987); Ireland,R. et al., J. Am. Chem. Soc. 110, 5768-5779 (1988); and Nakamura, H. etal., Proc. Nat. Acad. Sci. USA 86, 9672-9676 (1989)].

This invention is illustrated further by the following examples.

EXAMPLE 1 20-Deoxy-20-(2'-hydroxyethylthio)phorbol 12,13-Dibutyrate

One gram of sodium metal was dissolved in 50 mL methanol, and 44 μL ofthis solution was placed in a test tube. Then 2.63 grams distilled2-mercaptoethanol was dissolved in 50 mL acetonitrile, and 44 μL of thissolution was added to the test tube. Then 20 mg20-deoxy-20-chlorophorbol 12,13-dibutyrate were dissolved in 0.25 mLacetonitrile in a capped, nitrogen-flushed test tube. This lattersolution was then rapidly treated with the methoxide/mercaptoethanolsolution. An immediate precipitate formed. After 7 minutes, the reactionwas freed of solvent and treated with 1 mL water and 1 drop acetic acid.This residue was partitioned between water and ethyl acetate, followedby drying of the organic phase over sodium sulfate. Silica gelpreparative liquid chromatography using hexane/ethyl acetate mixturesyielded 9.5 mg of 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-dibutyrate, which could not be crystallized.

EXAMPLE 20 20-Deoxy-20-(2'-hydroxyethylthio)phorbol 12-Butyrate

Further preparative liquid chromatography of the reaction mixture fromExample 1 using hexane/ethyl acetate on silica gel yielded 5.7 mg20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-butyrate, the latter beingthe more polar compound. This compound could not be crystallized.

EXAMPLE 3 20-Deoxy-20-(2'-hydroxyethylthio)phorbol 12-Myristate13-Acetate

To 0.1 gram 20-deoxy-20-chlorophorbol 12-myristate 13-acetate in 1 mLacetonitrile was added 0.5 mL of a solution of 246 mg 2-mercaptoethanoland 436 mg 2,4,6-collidine in 10 mL acetonitrile, followed by 50 mgdiisopropylethylamine in 0.2 mL acetonitrile and 0.1 mL t-butyl methylether. Ten minutes later the reaction was treated with 0.107 mmolessodium methoxide and 0.225 mmoles 2-mercaptoethanol in 0.25 mL methanol.Five minutes later the proportion of sodium methoxide/mercaptoethanolwas doubled. After 10 minutes the reaction was stopped by addition of 2drops acetic acid and 0.5 mL water. The organics were extracted intoethyl acetate, washed once with water, and dried over sodium sulfate.After solvent removal, the crude residue was purified by preparativeliquid chromatography on silica gel using hexane/ethyl acetate (60:40).The product was 87 mg 20-deoxy-20-(2'-hydroxyethylthio)phorbol12-myristate 13-acetate in high purity. The compound did notcrystallize.

EXAMPLE 4 20-Deoxy-20-(2'-hydroxyethylthio)phorbol 12-Myristate

Twenty-five mg of 20-deoxy-20-chlorophorbol 12-myristate 13-acetate wasdissolved in 0.2 mL ethylene glycol and 0.2 mL acetonitrile. Thissolution was treated with 0.13 mL of a solution of 200 mg sodium metalin 20 mL ethylene glycol over a period of 40 minutes. The reaction waspartitioned between water and ethyl acetate, and the separated organicswere dried over sodium sulfate. After removal of the ethyl acetate, thecrude 20-deoxy-20-chlorophorbol 12-myristate was dissolved in 0.8 mLacetonitrile and treated with 0.18 mL of a solution of 0.31 mL2-mercaptoethanol, 0.5 mL acetonitrile, and 0.5 mL of 1% sodium inmethanol. After 40 minutes, an additional 0.04 mL of 1% sodium inmethanol was added. Ten minuted later the reaction was stopped with 1drop of acetic acid. After removal of the solvents in a stream ofnitrogen, the residue was partitioned between ethyl acetate and pH 8potassium phosphate. The organics were dried over sodium sulfate andfreed of solvent prior to preparative liquid chromatographicpurification using silica gel and hexane/ethyl acetate (45:55). Theproduct, 37 mg of20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-myristate,could not be crystallized.

EXAMPLE 5 20-Deoxy-20-[(2'-hydroxyethyl)methylamino]phorbol 12-Myristate13-Acetate

25 mg 20-deoxy-20-chlorophorbol 12-myristate 13-acetate was dissolved in0.4 mL acetonitrile. To this was added 0.1 mL of a solution of 29.6 mg2-(methylamino)ethanol in 1 mL acetonitrile. After 70 minutes andadditional 0.1 mL of the same amine solution was added. After anadditional 70 minutes, 0.2 mL more amine solution was added. After 6.6hours of total reaction time, the reaction was diluted with 4 mLmethylene chloride and subjected to preparative liquid chromatography onsilica gel using methylene chloride/methanol (92:8) followed byrepurification on silica gel using methylene chloride/methanol (96:4).The product, 20-deoxy-20-[(2'-hydroxyethyl)methylamino]phorbol12-myristate 13-acetate, 14 mg, could not be crystallized.

EXAMPLES 6, 7, and 8 20-Deoxy-20-fluorophorbol12,13-Bis[(2',4'-difluorophenyl)acetate], 12-beta,13-Bis[(2',4'-difluorophenyl)acetoxyl-4,9-dihydroxy-1,6(20),7-tigliatrien-3-oneand 12-beta,13-Bis[(2',4'-difluorophenyl)acetoxy]-4,9-dihydroxy-7-fluoro-1,6(20)-tigliadien-3-one

100 mg of phorbol 12,13-bis[(2',4'-difluorophenyl)acetate] weredissolved in 1.5 mL methylene chloride and the solution was set at 0° C.Then 26.2 mg diethylaminosulfur trifluoride in 0.5 mL methylene chloridewere added drop wise during 1 minute. After 40 minutes, 0.2 mL morediethylaminosulfur trifluoride was added. After 10 more minutes, thereaction was shaken with 2 mL pH 8 potassium phosphate buffer, afterwhich the organics were separated and dried over sodium sulfate. Thereaction was repeated twice more, and the combined reaction productswere freed of solvent, taken up in 10 mL ethyl acetate, and suckedthrough a funnel containing a layer of silica at the bottom, a layer ofsodium sulfate in the middle and sodium chloride at the top. Afterwashing the funnel contents with 50 mL ethyl acetate the combinedeluants were freed of solvent and repeatedly chromatographed on silicapreparative liquid chromatography columns using hexane/ethyl acetate(85:15) solvent mixtures. The products were 20-deoxy-20-fluorophorbol12,13-bis[(2',4'-difluorophenyl)acetate], 40 mg; 12-beta,13-bis[(2',4'-difluorophenyl)acetoxy]-4,9-dihydroxy-1,6(20),7-tigliatrien-3-one,25 mg; and 12-beta,13-bis[(2',4'-difluorophenyl)acetoxy]4,9-dihydroxy-7-fluoro-1,6(20)-tigliadien-3-one,40 mg; none of which could be crystallized.

EXAMPLE 9 4-Carboxy-6-(N-decanoylamino)indole

Five hundred-fifteen mg of 4-carbomethoxy-6-(N-decanoylamino)indole[prepared by the method of Wender et al., PNAS, 83, 4214-4218 (1986)]was dissolved in 60 mL of tetrahydrofuran. This solution was treatedwith 3 mL of a 1N KOH solution in water and also with 5 mL of methanol.The mixture was heated at 80° C. for 32 h. During this period another2.5 mL of 1N KOH was added in two portions. After cooling the mixturewas concentrated in vacuo. The mixture was then diluted with water andacidified with concentrated hydrochloric acid. The mixture was thenextracted with methylene chloride. The organic layers were dried oversodium sulfate and concentrated to afford 300 mg of4-carboxy-6-(N-decanoylamino)indole (60% yield), mp 248°-50° C.

EXAMPLE 10 4-Carboxy-6-(N-decanoylamino)indole N-Succinimidyl Ester

A suspension of 470 mg of 4-carboxy-6-(N-decanoylamino)indole and 427 mgof N-hydroxysuccinimide in 100 mL of acetonitrile, 10 mL of methylenechloride and 10 mL of tetrahydrofuran was prepared. This suspension wasstirred vigorously by a magnetic stirring bar as a solution of 540 mg ofdicyclohexylcarbodiimide in 20 mL of acetonitrile was slowly added overa period of 1 h. The mixture was stirred vigorously for 72 h. It wasthen concentrated in vacuo and diluted with ethyl acetate. The resultingmixture was filtered to remove the copious precipitate. The filtrate waswashed with water, dried over sodium sulfate and concentrated. Treatmentof the resulting mixture with hexane/ethyl acetate (50:50) followed byfiltration afforded 600 mg of 4-carboxy-6-(N-decanoylamino)indoleN-succinimidyl ester, mp 154°-5° C.

EXAMPLE 114-(N-2',3'-Dihydroxypropylcarboxamido)-6-(N-decanoylamino)indole

To a solution of 89 mg of 3-amino-1,2-propanediol in 20 mL oftetrahydrofuran was added 136 mg of4-carboxy-6-(N-decanoylamino)indoleN-succinimidyl ester. The solution was stirred for 48 h. Afterconcentration in vacuo the mixture was purified by preparative liquidchromatography using silica gel and methylene chloride/methanol (92:8).The product, 135 mg of4-(N-2',3'-dihydroxypropylcarboxamido)-6-(N-decanoylamino)indole, wasrecrystallized from methanol/methylene chloride, mp 139°-41° C.

EXAMPLE 12 4-(N-2'-Mercaptoethylcarboxamido)-6-(N-decanoylamino)indoleand 4-(S-2'-Aminoethylthiolcarbonyl)-6-(N-decanoylamino)indole

To a solution of 113 mg of triethylamine in 20 mL of tetrahydrofuran wasadded, first, 86 mg of 2-aminoethanethiol hydrochloride and, then, 139mg of 4-carboxy-6-(N-decanoylamino)indole N-succinimidyl ester. Thesolution was stirred for 48 h and then concentrated in vacuo.Purification by liquid chromatography using silica gel and methylenechloride/methanol (96:4) afforded two products. The earlier elutingproduct, 23 mg of4-(N-2'-mercaptoethylcarboxamido)-6-(N-decanoylamino)indole, decomposedat 200°-5° C. The later eluting product, 22 mg of4-(S-2'-aminoethylthiolcarbonyl)-6-(N-decanoylamino)indole, wasrecrystallized from methanol, mp 192°-3° C.

EXAMPLE 134-(2'-Hydroxymethylpiperidinocarbonyl)-6-(N-decanoylamino)indole

To a solution of 145 mg of 4-carboxy-6-(N-decanoylamino)indoleN-hydroxysuccinimidyl ester in 20 mL of tetrahydrofuran was added 103 mgor piperidinemethanol. The solution was heated at reflux for 5 days, atwhich time another 100 mg of piperidinemethanol was added and heatingcontinued for another day. The solution was then concentrated in vacuoand purified by liquid chromatography using silica gel and methylenechloride/isopropyl alcohol (93:7). In this manner 21 mg of4-(2'-hydroxymethylpiperidinocarbonyl)-6-(N-decanoylamino)indole wasobtained, mp 126°-129° C.

EXAMPLE 14 Phorbol 12-Myristate 13-Acetate 20-Methylcarbamate

One hundred milligrams of phorbol 12-myristate 13-acetate was dissolvedin 1 mL tetrahydrofuran. To this solution was added 11.5 μL of methylisocyanate, followed immediately by 20 μL of a 10% by weight solution ofdibutyltin dilaurate in tetrahydrofuran. After three hours an additional11.5 μL of methyl isocyanate was added. Sixteen hours later 30 μL of thereaction solution was applied to a silica gel TLC plate and developedwith hexanes/ethyl acetate (46:54). The band at R_(f) =0.4 was scrapedoff and the product was eluted from the silica with acetone. Removal ofthe solvent in a stream of nitrogen gave 2.8 mg of phorbol 12-myristate13-acetate 20-methylcarbamate as a glassy solid for spectroscopicanalysis and bioassay.

EXAMPLE 1512-beta-Myristoyloxy-13-acetoxy-4,9-dihydroxy-7-hydroxymethyl-1,6(20)-tigliadien-3-one

Ninety-eight mg of 20-deoxy-20-chlorophorbol 12-myristate 13-acetate wasdissolved in 0.5 mL tetrahydrofuran. To this was added 0.15 mL saturatedaqueous ammonium chloride, 0.15 mL 37% formalin solution, and 50 mg zincdust (less than 325 mesh). The reaction was capped and shaken vigorouslyfor 24 hours. At the end of this time the reaction was partitionedbetween pH 8 phosphate buffer and ethyl acetate. The ethyl acetate phasewas reduced to a volume of 2 mL under a stream of nitrogen and 0.125 mLwas applied to a silica gel TLC plate and developed with hexanes/ethylacetate (46:54). The band at R_(f) =0.45 was scraped off and the productwas eluted from the silica powder with acetone. Removal of the acetoneyielded 3.0 mg of12-beta-myristoyloxy-13-acetoxy-4,9-dihydroxy-7-hydroxymethyl-1,6(20)-tigliadien-3-oneas a glassy solid for spectroscopic analysis and bioassay.

EXAMPLE 16

Method: A stock solution of 300 pmoles of the standard inflammatorycompound phorbol 12-myristate 13-acetate per 5 μL acetone was prepared.This solution was used to prepare four-fold dilutions of20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-myristate 13-acetate,prepared as in Example 3, covering concentrations of the latter rangingfrom 4 to 64,000 pmoles per 5 μL. These solutions were used todemonstrate the anti-inflammatory activity of the latter compound byapplication of 5 μL to the insides of the right ears of mice, followedby the observation of ear inflammation/erythema at intervals from 1 to48 hours after application. Inhibition of the phorbol 12-myristate13-acetate induced inflammation was observed at the medium and higherconcentrations of the inhibitor.

In a like manner, the anti-inflammatory activities of the followingcompounds are demonstrated; lower doses produce shorter periods ofinflammation/erythema and higher doses produce longer periods of, and inmost cases complete, inhibition during the entire assay period.

A. Diterpenoid-type phorboids:

(i) phorbol 12-myristate;

(ii) phorbol 12,13-diacetate;

(iii) 20-deoxy-20-chlorophorbol 12-myristate 13-acetate;

(iv) 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12,13-dibutyrate;

(v) 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-myristate 13-acetate;

(vi) 20-deoxy-20-[(2'-hydroxyethyl)methylamino]phorbol 12-myristate13-acetate;

(vii) 20-deoxy-20-fluorophorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(viii) 12-beta,13-bis[(2',4'-difluorophenyl)acetoxy]-4,9-dihydroxy-1,6(20),7-tigliatrien-3-one;

(ix) 12-beta,13-bis[(2',4'-difluorophenyl)acetoxy]-4,9-dihydroxy-7-fluoro-1,6(20)-tigliadien-3-one;

(x) 12-beta-myristoyloxy13-acetoxy-4,9-dihydroxy-7-hydroxymethyl-1,6(20)-tigliadien-3-one;

(xi) phorbol 12-myristate 13-acetate 20-methylcarbamate;

(xii) 6-deshydroxymethyl-6-carboxyphorbol 12,13-didecanoate;

(xiii) 6-deshydroxymethyl-6-carboxyphorbol 12-octyldimethylsilylacetate13-(2',4'-difluorophenyl)acetate;

(xiv) 6-deshydroxymethyl-6-carboxyresiniferonol9,13,14-orthophenylacetate;

(xv) 20-deshydroxy-20-carboxyingenol 3-benzoate;

(xvi) 6-deshydroxy-20-carboxymethylidenephorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(xvii) 20-deoxy-20-dimethylphosphinylphorbol 12-myristate 13-acetate;

(xviii) 20-deoxy-20-dimethylphosphonylphorbol 12-myristate 13-acetate;

(xix) 20-deoxy-20-cyanophorbol 12-myristate 13-acetate;

(xx) phorbol 12-[4'-(9",10"-dihydrophenanthrene-2")butyrate];

(xxi) phorbol 12-(pentafluorophenyl)acetate;

(xxii) 20-deoxy-20-azidophorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(xxiii) 20-deoxy-20-mercaptophorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(xxiv) 20-deoxy-20-hydroximinophorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(xxv) 6-deshydroxymethyl-6-carboxyphorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(xxvi) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(xxvii) 20-deoxyphorbol12,13-bis[(2',4'-diflourophenyl)acetate]20-sulfonic acid; and

(xxviii) 20-deoxy-20-azidophorbol12,13-bis[(2',4'-difluorophenyl)acetate].

B. Indolactam-type phorboids:

(i)rac-14-O-(N-methyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethylindolactamV, methanesulfonate salt;

(ii) 14-O-(N-methyl)carbamoyl-7-octyl-(9S,12S)-indolactam V;

(iii)9-deshydroxymethyl-9-[N-(2',3'dihydroxypropyl)]carboxamidoindolactam V;

(iv) 9-deshydroxy-9-(2'-hydroxyethylthio)indolactam V;

(v) 9-deshydroxy-9-(3'-hydroxypropylthio)-7-octyl-(9S,12S)-indolactam V;

(vi) 9-deshydroxymethyl-9-carboxyindolactam V, triethylamine salt;

(vii) 9-deshydroxy-9-(2'-hydroxyethylthio)-7-octyl-(9S,12S)-indolactamV;

(viii) 7-octyl-(9S,12S)-indolactam V 14-O-(N'-methylcarbamate);

(ix) 1-(2'-triphenylphosphonium)ethyl-7-octyl-(9S,12S)-indolactam V14-O-(N'-methylcarbamate);

(x) 9-deshydroxymethyl-9-carboxyindolactam V; and

(xi) 14-deoxy-14-(3'-hydroxypropylthio)-7-octyl-(9S,12S)-indolactam V.

EXAMPLE 17 9-Deshydroxymethyl-9-carboxyindolactam V

A solution of 60 g of 4-nitrogramine [J. B. Hester, J. Org. Chem., 29,1158 (1964)] and 54 g of ethyl nitroacetate in 1.2 L of chlorobenzenewas heated at 100° C. for 1.5 h. After cooling the mixture was filtered,washed with cold methylene chloride and dried in vacuo to afford 58.9 gof ethyl 3-(4'-nitroindol-2'-yl)-2-nitropropionate as a yellow solid: mp159°-160.5° C. Another 11 g may be recovered from the filtrate bydilution with hexane, preparative liquid chromatography [silica;methylene chloride/ethyl acetate (90:10)] and recrystallization frommethanol. The structure was confirmed by NMR.

To a solution of 7.29 g of ethyl3-(4'-nitroindol-2'-yl)-2-nitropropionate in 100 mL of tetrahydrofuranand 100 mL of ethanol was added 721 mg of 10% Pd on carbon. Theresulting mixture was shaken in a Parr apparatus under about 50 psihydrogen. After 70 min the mixture was filtered through celite andwashed with ethanol. The filtrate was concentrated in vacuo. Afterpurification by preparative liquid chromatography [silica;hexane/tetrahydrofuran (60:40)] 5.5 g of ethyl3-(4'-aminoindol-2'-yl)-2-nitropropionate was obtained as an off-whitesolid, mp 110°-112° C. The structure was confirmed by NMR.

To a mixture prepared by treatment of 8.6 g of the sodium salt of3-methyl-2-oxobutanoic acid in 40 mL of dimethylformamide with 63 mmoleof hydrogen chloride in 17 mL of dimethylformamide was added 10 g ofethyl 3-(4'-aminoindol-2'-yl)-2-nitropropionate in 45 mL ofN,N-dimethylformamide. After the resulting mixture had been cooled in anice water bath, a solution of 6 g of sodium cyanoborohydride in 45 mL ofN,N-dimethylformamide was added over 15 min. After the addition wascomplete, the mixture was allowed to warm to room temperature over aperiod of 30 min, at which time 200 mL of water was added and themixture acidified with 2N hydrochloric acid. This solution was extractedwith ethyl acetate. The organic layers were combined, washed with brine,dried over sodium sulfate and concentrated in vacuo and at a temperatureonly slightly above ambient to afford a crude mixture containingN-[4-[3-(2'-nitro-2'-ethoxycarbonyl)ethyl]indolyl]valine.

To a cooled solution of this crude residue and 7.2 g ofN-hydroxysuccinimide in 250 mL of acetonitrile was added 15.5 g ofdicyclohexylcarbodiimide in 35 mL of acetonitrile. After 40 min, 2.7 mLof glacial acetic acid was added. After another 20 min the mixture wasfiltered and washed with ethyl acetate. The filtrates were washed withwater and brine, dried over sodium sulfate and concentrated in vacuo toafford crude N-[4-[3-(2'-nitro-2'-ethoxycarbonyl)ethyl]indolyl]valineN-succinimidyl ester. After combination with another batch andpurification by preparative liquid chromatography [silica;hexane/tetrahydrofuran (57:43)], 24.4 g (91% yield) of the ester wasobtained as a gum. The structure was confirmed by NMR and mass spectralanalysis.

To a solution of 3.75 g ofN-[4-[B-(2'-nitro-2'-ethoxycarbonyl)ethyl]indolyl]valine N-succinimidylester in 200 mL of methanol was added 11.5 mL of a slurry of Raneynickel. This mixture was shaken on a Parr apparatus under about 50 psiof hydrogen for 35 min. The supernatant was removed by decantation andconcentrated in vacuo. Several such crude mixtures were combined andpurified by preparative liquid chromatography [silica;hexane/tetrahydrofuran (60:40)] to afford 3.23 g (31% yield) ofN-desmethyl-9-deshydroxymethyl-9-ethoxycarbonylindolactam V, mp 195°-5°C. (dec), and 2.13 g (20.5% yield) ofN-desmethyl-9-deshydroxymethyl-9-ethoxycarbonyl-epi-indolactam V, mp206°-8° C. (dec). The structures of these compounds were confirmed byNMR and mass spectral analysis and by conversion to the known indolactamV and epi-indolactam V respectively.

To a solution of 204 mg ofN-desmethyl-9-deshydroxymethyl-9-ethoxycarbonylindolactam V in 20 mL ofacetonitrile containing 1.5 mL of water was added 400 μL of 37% aqueousformaldehyde. After 20 min, 182 mg of sodium cyanoborohydride was added.After the mixture had stirred at room temperature for 3 hours, phosphatebuffer (pH 2) was added. The mixture was then concentrated in vacuobefore re-diluting with water and extracting with ethyl acetate. Theorganic layer was dried over sodium sulfate, filtered and concentratedin vacuo. The resulting solid was purified by preparative liquidchromatography [silica; hexane/tetrahydrofuran (60:40)] to afford 167 mgof 9-deshydroxymethyl-9-ethoxycarbonylindolactam V. The structure wasconfirmed by NMR and mass spectral analysis.

To a solution of 160 mg of9-deshydroxymethyl-9-ethoxycarbonylindolactamV in 23 mL of methanol was added 2.3 mL of 2N sodium hydroxide. Afterone hour 2N hydrochloric acid was added until the mixture was acidicwhereupon it was concentrated to a small volume in vacuo. The residuewas then diluted with water and extracted with ethyl acetate. Theorganic layer was dried over sodium sulfate, filtered and concentratedin vacuo to afford 137 mg of 9-deshydroxymethyl-9-carboxyindolactam V asa white solid.

EXAMPLE 18 9-Deshydroxymethyl-9-carboxy-epi-indolactam V

A solution of 10 mg ofN-desmethyl-9-deshydroxymethyl-9-ethoxycarbonyl-epi-indolactam V in 1 mLof acetonitrile containing 90 μL of water was added 20 μL of 37% aqueousformaldehyde. After 15 min 9 mg of sodium cyanoborohydride was added.After the mixture had stirred at room temperature for 3.5 hours,phosphate buffer (pH 2) was added and the mixture further diluted withwater and extracted with ethyl acetate. The organic layer was dried oversodium sulfate, filtered and concentrated in vacuo to afford crude9-deshydroxymethyl-9-ethoxycarbonyl-epi-indolactam V.

To a solution of 9-deshydroxymethhyl-9-ethoxycarbonyl-epi-indolactam Vin 2 mL of methanol was added 150 μL of 2N sodium hydroxide. After 50min 2N hydrochloric acid was added until the mixture was acidicwhereupon it was then diluted with water and extracted with ethylacetate. The organic layer was dried over sodium sulfate, filtered andconcentrated in vacuo to afford a residue of9-deshydroxymethyl-9-carboxy-epi-indolactam V. The structure wasconfirmed by conversion to9-deshydroxymethyl-9-methoxycarbonyl-epi-indolactam V by treatment witha solution of diazomethane in ether.

EXAMPLE 19 9-Deshydroxymethyl-9-carboxyindolactam V N-Succinimidyl Ester

To a solution of 137 mg of 9-deshydroxymethyl-9-carboxyindolactam V and66 mg of N-hydroxysuccinimide in 7 mL of acetonitrile was added 119 mgof dicyclohexylcarbodiimide in 3 mL of acetonitrile. After about onehour the reaction mixture was filtered and then concentrated in vacuo.Purification by preparative liquid chromatography [silica;hexane/tetrahydrofuran (70:30)] afforded 136 mg of9-deshydroxymethyl-9-carboxyindolactam V N-succinimidyl ester.

EXAMPLE 209-Deshydroxymethyl-9-[N-(2',3'-dihydroxy)propyl]carboxamidoindolactam V

To 11 mg of 3-amino-1,2-propandiol and 1 mg of 4-dimethylaminopyridinewas added 25 mg of 9-deshydroxymethyl-9-carboxyindolactam VN-succinimidyl ester in 250 μL of methylene chloride and 0.4 mLtetrahydrofuran. After 5 h the mixture was concentrated in vacuo. Afterpreparative liquid chromatography [silica; methylene chloride/isopropylalcohol (85:15); followed by ODS silica; acetonitrile/water (33:67)],10.5 mg of9-deshydroxymethyl-9-[N-(2',3'-dihydroxy)propyl]carboxamidoindolactam Vand 3.4 mg of9-deshydroxymethyl-9-[N-(2',3'-dihydroxy)propyl]carboxamido-epi-indolactamV were obtained. The structures of these compounds were confirmed bymass spectral analysis.

EXAMPLE 219-Deshydroxymethyl-9-(2',3'-dihydroxy)propyloxycarbonylindolactam V

To 30 mg of glycerol and 1 mg of 4-dimethylaminopyridine was added 25 mgof 9-deshydroxymethyl-9-carboxyindolactam V N-succinimidyl ester in 250μL of methylene chloride and 0.4 mL tetrahydrofuran. After 20 h themixture was concentrated in vacuo. After preparative liquidchromatography [silica; methylene chloride/isopropyl alcohol (91:9);followed by ODS silica; acetonitrile/water (36:64)] 13.3 mg of9-deshydroxymethyl-9-(2',3'-dihydroxy)propyloxycarbonylindolactam V and1.2 mg of9-deshydroxymethyl-9-(2',3'-dihydroxy)propyloxycarbonyl-epi-indolactam Vwere obtained. The structures of these compounds were confirmed by massspectral analysis.

EXAMPLE 22 9-Deshydroxymethyl-9-[N-(2'-glucosyl)]carboxamidoindolactam V

To 29 mg of 2-glucosamine hydrochloride, 19 mg of triethylamine and 1 mgof 4-dimethylaminopyridine was added 25 mg of9-deshydroxymethyl-9-carboxyindolactam V N-succinimidyl ester in 250 μLof methylene chloride and 0.4 mL tetrahydrofuran. After 7.5 h themixture was concentrated in vacuo. After preparative liquidchromatography [silica; methylene chloride/methanol (80:20); followed byODS silica; acetonitrile/water (1.2% triethylamine) (21:79)], 12.4 mg of9-deshydroxymethyl-9-[N-(2'-glucosyl)]carboxamidoindolactam V and 2.4 mgof 9-deshydroxymethyl-9-[N-(2'-glucosyl)]carboxamido-epi-indolactam Vwere obtained.

EXAMPLE 23

In a similar manner the following compounds are prepared:

(i) 9-deshydroxymethyl-9-[N-(2'-carboxy)ethyl]carboxamidoindolactam V;

(ii) 9-deshydroxymethyl-9-[N-(2'-hydroxy)ethyl]carboxamidoindolactam V;

(iii) 9-deshydroxymethyl-9-(2'-hydroxy)ethylthiocarbonylindolactam V;

(iv) 9-deshydroxymethyl-9-(2'-hydroxy)ethoxycarbonylindolactam V;

(v)9-deshydroxymethyl-9-[N-(2',3'-dihydroxy)propyl]carboxamido-7-octylindolactamV;

(vi)9-deshydroxymethyl-9-[N-(2'-carboxy)ethyl]carboxamido-7-octylindolactamV;

(vii)9-deshydroxymethyl-9-[N-(2'-hydroxy)ethyl]carboxamido-7-octylindolactamV;

(viii)9-deshydroxymethyl-9-(2'-hydroxy)ethylthiocarbonyl-7-octylindolactam V;

(ix) 9-deshydroxymethyl-9-(2'-hydroxy)ethoxycarbonyl-7-octylindolactamV;

(x)9-deshydroxymethyl-9-[N-(2',3'-dihydroxy)propyl]carboxamido-7-octyl-epi-indolactamV;

(xi)9-deshydroxymethyl-9-[N-(2'-carboxy)ethyl]carboxamido-7-octyl-epi-indolactamV;

(xii)9-deshydroxymethyl-9-[N-(2'-hydroxy)ethyl]carboxamido-7-octyl-epi-indolactamV;

(xiii)9-deshydroxymethyl-9-(2'-hydroxy)ethylthiocarbonyl-7-octyl-epi-indolactamV;

(xiv)9-deshydroxymethyl-9-(2'-hydroxy)ethoxycarbonyl-7-octyl-epi-indolactamV;

(xv)9-deshydroxymethyl-9-[N-(2',3'-dihydroxy)propyl]carboxamido-6,7-tetramethyleneindolactamV;

(xvi)9-deshydroxymethyl-9-[N-(2'-carboxy)ethyl]carboxamido-6,7-tetramethyleneindolactamV;

(xvii)9-deshydroxymethyl-9-[N-(2'-hydroxy)ethyl]carboxamido-6,7-tetramethyleneindolactamV;

(xviii)9-deshydroxymethyl-9-(2'-hydroxy)ethylthiocarbonyl-6,7-tetramethyleneindolactamV;

(xix)9-deshydroxymethyl-9-(2'-hydroxy)ethoxycarbonyl-6,7-tetramethyleneindolactamV;

(xx)9-deshydroxymethyl-9-[N-(2',3'-dihydroxy)propyl]carboxamido-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxi)9-deshydroxymethyl-9-[N-(2'-carboxy)ethyl]carboxamido-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxii)9-deshydroxymethyl-9-[N-(2'-hydroxy)ethyl]carboxamido-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxiii)9-deshydroxymethyl-9-(2'-hydroxy)ethylthiocarbonyl-7-octyl-12-des-iso-propyl-12-benzylindolactamV; and

(xxiv)9-deshydroxymethyl-9-(2'-hydroxy)ethoxycarbonyl-7-octyl-12-des-iso-propyl-12-benzylindolactamV.

EXAMPLE 2414-O-[N-(S)-(1'-Naphthyl)ethyl]carbamoyl-7-octyl-(9S,12S)-indolactam Vand 14-O-[N-(S)-(1'-Naphthyl)ethyl]carbamoyl-7-octyl-(9R,12R)-indolactamV

To a solution of 139 mg of racemic 7-octylindolactam V [prepared as inK. Irie, et al. Agric. Biol. Chem., 50, 2679 (1986)] in 15 mL ofanhydrous tetrahydrofuran was added 56 mg of dibutyltin dilaurate and 48mg of 4-dimethylaminopyridine in 3 mL of tetrahydrofuran. To thissolution was added 445 mg of (S)-1-(1-napthyl)ethyl isocyanate in twoportions over a two day period. The mixture was then concentrated invacuo and the residue partitioned between water and ethyl acetate. Theorganic layer was dried over sodium sulfate and concentrated in vacuo.After preparative liquid chromatography [silica; hexane/tetrahydrofuran(75:25)], 200 mg of a mixture of diastereomers was obtained. Repetitivechromatography of this mixture [wet silica; hexane/wet ethyl acetate(65:35)] afforded 87 mg of pure14-O-[N-(S)-(1'-naphthyl)ethyl]carbamoyl-7-octyl-(9S,12S)indolactam Vand 96 mg of pure14-O-[N-(S)-(1'-naphthyl)ethyl]carbamoyl-7-octyl-(9R,12R)-indolactam V.The structures of these compounds were confirmed by reduction to theknown (-)-7-octylindolactam V and (+)-7-octylindolactam V respectively.

EXAMPLE 25 14-O-[N-(R)-(1'-Naphthyl)ethyl]carbamoyl-(9S,12S)-indolactamV and 14-O-[N-(R)-(1'-Naphthyl)ethyl]carbamoyl-(9R,12R)-indolactam V

To a solution of 2.3 g of racemic indolactam V in 80 mL of anhydroustetrahydrofuran with 569 mg of dibutyltin dilaurate and 560 mg of4-dimethylaminopyridine was added 2.4 g of (R)-1-(1-napthyl)ethylisocyanate. After stirring at room temperature for 24 h, the mixture wasconcentrated in vacuo and the residue partitioned between water andethyl acetate. The organic layer was dried over sodium sulfate andconcentrated in vacuo. After preparative liquid chromatography [silica;hexane/tetrahydrofuran (55:45)], 3.65 g of a mixture of diastereomerswas obtained. Repetitive chromatography of this mixture [wet silica;hexane/wet ethyl acetate (65:35)] afforded 1.92 g of14-O-[N-(R)-(1'-naphthyl)ethyl]carbamoyl-(9R,12R)-indolactam V and 1.77g of 14-O-[N-(R)-(1'-naphthyl)ethyl]carbamoyl-(9S,12S)-indolactam V. Thestructures of these compounds were confirmed by NMR, by comparison witha sample of 14-O-[N-(R)-(1'-naphthyl)ethyl]carbamoyl-(9S,12S)-indolactamV prepared from authentic (-)-indolactam V, and by reduction to theknown (+)-indolactam V and (-)indolactam V respectively.

EXAMPLE 26 14-O-(N-Methyl)carbamoyl-7-octyl-(9S,12S)-indolactam V

A solution of 3 mg of dibutyltin dilaurate and 2 mg of4-dimethylaminopyridine in 0.5 mL of anhydrous tetrahydrofuran was addedto 5 mg of (-)-7-octylindolactam V. Then 5 μL of methylisocyanate wasadded. After four hours at room temperature the mixture was concentratedunder nitrogen and purified by preparative liquid chromatography[silica; hexane/tetrahydrofuran (70:30)] to afford 5 mg of14-O-(N-methyl)carbamoyl-7-octyl-(9S,12S)-indolactam V.

EXAMPLE 27 rac-14-O-(N-Methyl)carbamoylindolactam V

To a solution of 105 mg of racemic indolactam V, 41 mg of4-dimethylaminopyridine, and 57 mg of dibutyltin dilaurate in 18 mL ofanhydrous tetrahydrofuran was added 150 μL of methylisocyanate in twoportions over 1.5 h. One hour after the final addition the mixture wasconcentrated in vacuo and the residue purified by recrystallization fromtetrahydrofuran with hexane to afford 100 mg ofrac-14-O-(N-methyl)carbamoylindolactam V, mp 184°-185° C.

EXAMPLE 28

In a similar manner the following compounds are prepared:

(i) 14-O-(N-ethyl)carbamoylindolactam V;

(ii) 14-O-(N-methyl)thiocarbamoylindolactam V;

(iii) 14-O-(N-benzyl)carbamoylindolactam V;

(iv) 14-O-(N-ethyl)carbamoyl-7-octylindolactam V;

(v) 14-O-(N-methyl)thiocarbamoyl-7-octylindolactam V;

(vi) 14-O-(N-benzyl)carbamoyl-7-octylindolactam V;

(vii) 14-O-(N-methyl)carbamoyl-7-octyl-epi-indolactam V;

(viii) 14-O-(N-ethyl)carbamoyl-7-octyl-epi-indolactam V;

(ix) 14-O-(N-methyl)thiocarbamoyl-7-octyl-epi-indolactam V;

(x) 14-O-(N-benzyl)carbamoyl-7-octyl-epi-indolactam V;

(xi) 14-O-(N-methyl)carbamoyl-6,7-tetramethyleneindolactam V;

(xii) 14-O-(N-ethyl)carbamoyl-6,7-tetramethyleneindolactam V;

(xiii) 14-O-(N-methyl)thiocarbamoyl-6,7-tetramethyleneindolactam V;

(xiv) 14-O-(N-benzyl)carbamoyl-6,7-tetramethyleneindolactam V;

(xv)14-O-(N-methyl)carbamoyl-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xvi)14-O-(N-ethyl)carbamoyl-7-octyl-12-des-iso-propyl-12-benzylindolactam V;

(xvii)14-O-(N-methyl)thiocarbamoyl-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xviii)14-O-(N-benzyl)carbamoyl-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xix) 14-O-(N-ethyl)carbamoylteleocidin B;

(xx) 14-O-(N-methyl)thiocarbamoylteleocidin B; and

(xxi) 14-O-(N-benzyl)carbamoylteleocidin B.

EXAMPLE 29 rac-14-O-(N-Methyl)carbamoyl-1-N-diphenylphosphorylindolactamV

To 12 mg of rac-14-O-(N-methyl)carbamoylindolactam V in 500 μL ofanhydrous tetrahydrofuran was added approximately 2 mg of sodium hydride(50% dispersion in oil) followed by 30 μL of diphenyl chlorophosphate intwo portions. After 2-3 hours, thin layer chromatographic analysis[silica; methylene chloride/methanol (95:5)] showed that the startingrac-14-O-(N-methyl)carbamoylindolactam V (R_(f) =0.36) had beenconverted torac-14-O-(N-methyl)carbamoyl-1-N-diphenylphosphorylindolactam V withR_(f) =0.43.

EXAMPLE 30rac-14-O-(N-Methyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethylindolactamV, Methanesulfonate salt

To 16 mg of rac-14-O-(N-methyl)carbamoylindolactam V in 0.75 mL ofN,N-dimethylformamide in an ice-water bath was added 8 mg of sodiumhydride (60% dispersion in oil). After about 10 min this solution wasadded to 20 mg of 2-methanesulfonyloxyethyltriphenylphosphonium bromide(prepared from 2-hydroxyethyltriphenylphosphonium bromide). After 1 hthis mixture was concentrated in vacuo. The residue was partitionedbetween ethyl acetate and phosphate buffer (pH 2). The organic layer wasdried over sodium sulfate and concentrated in vacuo. The crude mixturewas taken up in methanol, treated with a small amount of methanesulfonicacid in methanol and reconcentrated. Thin layer chromatographic analysis[silica; methylene chloride/methanol (90:10)] showed that the startingrac-14-O-(N-methyl)carbamoylindolactam V (R_(f) =0.63) had beenconverted torac-14-O-(N-methyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethylindolactamV, methanesulfonate salt, with R_(f) =0.57.

EXAMPLE 31

In a similar manner the following compounds are prepared;

(i)14-O-(N-methyl)thiocarbamoyl-1-N-(2'-triphenylphosphonium)ethylindolactamV, methanesulfonate salt;

(ii)14-O-(N-benzyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethylindolactam V,methanesulfonate salt;

(iii)14-O-(N-methyl)thiocarbamoyl-1-N-(2'-triphenylphosphonium)ethyl-epi-indolactamV, methanesulfonate salt;

(iv)14-O-(N-benzyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethyl-epi-indolactamV, methanesulfonate salt;

(v)14-O-(N-methyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethyl-epi-indolactamV, methanesulfonate salt;

(vi)14-O-(N-methyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethyl-12-des-iso-propyl-12-benzylindolactamV, methanesulfonate salt;

(vii)14-O-(N-methyl)thiocarbamoyl-1-N-(2'-triphenylphosphonium)ethyl-12-des-iso-propyl-12-benzylindolactamV, methanesulfonate salt; and

(viii)14-O-(N-benzyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethyl-12-des-iso-propyl-12-benzylindolactamV, methanesulfonate salt.

EXAMPLE 32rac-14-O-(N-Methyl)carbamoyl-1-N-trimethylsilylmethylindolactam V

To 16 mg of rac-14-O-(N-methyl)carbamoylindolactam V in 0.75 m L ofN,N-dimethylformamide in an ice-water bath was added 8 mg of sodiumhydride (60% dispersion in oil). After about 10 min, 20 μL ofbromomethyltrimethylsilane was added. After 1 h this mixture wasconcentrated in vacuo and the residue partitioned between ethyl acetateand phosphate buffer (pH 8). The organic layer was dried over sodiumsulfate and concentrated in vacuo. Thin layer chromatographic analysis[silica; methylene chloride/methanol (95:5)] showed that the startingrac-14-O-(N-methyl)carbamoylindolactam V (R_(f) =0.36) had beenconverted torac-14-O-(N-methyl)carbamoyl-1-N-trimethylsilylmethylindolactam V withR_(f) =0.59.

EXAMPLE 33

In a similar manner the following compounds are prepared:

(i) 14-O-(N-methyl)thiocarbamoyl-1-N-trimethylsilylmethylindolactam V;

(ii) 14-O-(N-benzyl)carbamoyl-1-N-trimethylsilylmethylindolactam V;

(iii) 14-O-(N-methyl)carbamoyl-1-N-trimethylsilylmethyl-epi-indolactamV;

(iv)14-O-(N-methyl)thiocarbamoyl-1-N-trimethylsilylmethyl-epi-indolactam V;

(v) 14-O-(N-benzyl)carbamoyl-1-N-trimethylsilylmethyl-epi-indolactam V;

(vi)14-O-(N-methyl)carbamoyl-1-N-trimethylsilylmethyl-12-des-iso-propyl-12-benzylindolactamV;

(vii)14-O-(N-methyl)thiocarbamoyl-1-N-trimethylsilylmethyl-12-des-iso-propyl-12-benzylindolactamV; and

(viii)14-O-(N-benzyl)carbamoyl-1-N-trimethylsilylmethyl-12-des-iso-propyl-12-benzylindolactamV.

EXAMPLE 3414-O-[(Diisopropylamino)methoxy]phosphinyl-7-octyl-(9S,12S)-indolactam V

To 5 mg of (-)-7-octylindolactam V in 180 μL of anhydrous methylenechloride was added 15 μL of diisopropylethylamine followed by 7 μL ofN,N-diisopropylmethylphosphoramidic chloride. After 0.5 h, thin layerchromatographic analysis [silica; hexane/ethyl acetate (45:55)] showedthat the starting (-)-7-octylindolactam V (R_(f) =0.17) had beenconverted to14-O-[(diisopropylamino)methoxy]phosphinyl-7-octyl-(9S,12S)-indolactam Vwith R_(f) =0.72.

EXAMPLE 35 14-O-(Dimethyl)thiophosphoryl-7-octyl-(9S,12S)-indolactam V

To 5 mg of (-)-7-octylindolactam V in 200 μL of anhydrous methylenechloride and containing 5 μL of pyridine was added 14 μL of dimethylchlorothiophosphate and approximately 10 mg of 4-dimethylaminopyridine.After 2 h, thin layer chromatographic analysis [silica; methylenechloride/methanol (95:5)] showed that the starting (-)-7-octylindolactamV (R_(f) =0.32) had been converted to14-O-(dimethyl)thiophosphoryl-7-octyl-(9S,12S)-indolactam V with R_(f)=0.36.

EXAMPLE 36

In a similar manner the following compounds are prepared:

(i) 14-O-(dimethyl)phosphorylindolactam V;

(ii) 14-O-(tetramethyl)phosphorodiamidylindolactam V;

(iii) 14-O-(diethyl)phosphonylindolactam V;

(iv) 14-O-[bis(2',2',2'-trichloroethyl)]phosphorylindolactam V;

(v) 14-O-(dimethyl)thiophosphorylindolactam V;

(vi) 14-O-(dimethyl)phosphoryl-7-octylindolactam V;

(vii) 14-O-(tetramethyl)phosphorodiamidyl-7-octylindolactam V;

(viii) 14-O-(diethyl)phosphonyl-7-octylindolactam V;

(ix) 14-O-[bis(2',2',2'-trichloroethyl)]phosphoryl-7-octylindolactam V;

(x) 14-O-(dimethyl)thiophosphoryl-7-octyl-epi-indolactam V;

(xi) 14-O-(dimethyl)phosphoryl-7-octyl-epi-indolactam V;

(xii) 14-O-(tetramethyl)phosphorodiamidyl-7-octyl-epi-indolactam V;

(xiii) 14-O-(diethyl)phosphonyl-7-octyl-epi-indolactam V;

(xiv)14-O-[bis(2',2',2'-trichloroethyl)]phosphoryl-7-octyl-epi-indolactam V;

(xv) 14-O-(dimethyl)phosphoryl-6,7-tetramethyleneindolactam V;

(xvi) 14-O-(tetramethyl)phosphorodiamidyl-6,7-tetramethyleneindolactamV;

(xvii) 14-O-(diethyl)phosphonyl-6,7-tetramethyleneindolactam V;

(xviii)14-O-[bis(2',2',2'-trichloroethyl)]phosphoryl-6,7-tetramethyleneindolactamV;

(xix) 14-O-(dimethyl)thiophosphoryl-6,7-tetramethyleneindolactam V;

(xx)14-O-(dimethyl)thiophosphoryl-7-octylo12-des-iso-propyl-12-benzylindolactamV;

(xxi)14-O-(dimethyl)phosphoryl-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxii) 14-O-(tetramethyl)phosphorodiamidyl-7-octylindolactam V;

(xxiii)14-O-(diethyl)phosphonyl-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxiv)14-O-[bis(2',2',2'-trichloroethyl)]phosphoryl-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxv) 14-O-(dimethyl)phosphorylteleocidin B;

(xxvi) 14-O-(tetramethyl)phosphorodiamidylteleocidin B;

(xvii) 14-O-(diethyl)phosphonylteleocidin B;

(xxviii) 14-O-[bis(2'2',2'-trichloroethyl)]phosphorylteleocidin B; and

(xxix) 14-O-(dimethyl)thiophosphorylteleocidin B.

EXAMPLE 3714-Deoxy-14-(3'-hydroxy)propylthio-7-octyl-(9S,12S)-indolactam V

To a solution of 6 mg of 3-mercapto-1-propanol in 150 μL of methanolcontaining 1.3 mg of sodium methoxide was added approximately 6 mg of14-O-methanesulfonyl-7-octyl-(9S,12S)-indolactam V (prepared from(-)-7-octylindolactam V) in 300 μL of acetonitrile. After 20 h thismixture was diluted with ethyl acetate and washed twice with phosphatebuffers (pH 2 and pH 8). After drying over sodium sulfate, the organiclayer was concentrated under a nitrogen stream. Thin layerchromatographic analysis [silica; methylene chloride/methanol (96:4)]showed that the starting14-O-methanesulfonyl-7-octyl-(9S,12S)-indolactam V (R_(f) =0.67) hadbeen converted to 14-deoxy-14-(3'-hydroxy)propylthio-7-octyl-(9S,12S)-indolactam V with R_(f) =0.22.

EXAMPLE 38

In a similar manner the following compounds are prepared:

(i) 14-deoxy-14-butylthio-7-octylindolactam V;

(ii) 14-deoxy-14-(2'-hydroxy-1'-methyl)ethylthio-7-octylindolactam V;

(iii) 14-deoxy-14-(2'-carboxy)ethylthio-7-octylindolactam V;

(iv) 14-deoxy-14-(2'-amino)ethylthio-7-octylindolactam V;

(v) 14-deoxy-14-(3'-hydroxymethyl)phenylthio-7-octylindolactam V;

(vi) 14-deoxy-14-propylthio-7-octyl-epi-indolactam V;

(vii) 14-deoxy-14-(2'-hydroxy-1'-methyl)ethylthio-7-octyl-epi-indolactamV;

(viii) 14-deoxy-14-(2'-carboxy)ethylthio-7-octyl-epi-indolactam V;

(ix) 14-deoxy-14-(2'-amino)ethylthio-7-octyl-epi-indolactam V;

(x) 14-deoxy-14-(3'-hydroxymethyl)phenylthio-7-octyl-epi-indolactam V;

(xi) 14-deoxy-14-(2'-hydroxy)ethylthio-7-octyl-epi-indolactam V;

(xii) 14-deoxy-14-propylthio-6,7-tetramethyleneindolactam V;

(xiii)14-deoxy-14-(2'-hydroxy-1'-methyl)ethylthio-6,7-tetramethyleneindolactamV;

(xiv) 14-deoxy-14-(2'-carboxy)ethylthio-6,7-tetramethyleneindolactam V;

(xv) 14-deoxy-14-(2'-amino)ethylthio-6,7-tetramethyleneindolactam V;

(xvi)14-deoxy-14-(3'-hydroxymethyl)phenylthio-6,7-tetramethyleneindolactam V;

(xvii) 14-deoxy-14-(2'-hydroxy)ethylthio-6,7-tetramethyleneindolactam V;

(xviii)14-deoxy-14-propylthio-7-octyl-12-des-iso-propyl-12-benzylindolactam V;

(xix)14-deoxy-14-(2'-hydroxy-1'-methyl)ethylthio-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xx)14-deoxy-14-(2'-carboxy)ethylthio-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxi)14-deoxy-14-(2'-amino)ethylthio-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxii)14-deoxy-14-(3'-hydroxymethyl)phenylthio-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxiii)14-deoxy-14-(2'-hydroxy)ethylthio-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(xxiv) 14-deoxy-14-propylthioteleocidin B;

(xxv) 14-deoxy-14-(2'-hydroxy-1'-methyl)ethylthioteleocidin B;

(xxvi) 14-deoxy-14-(2'-carboxy)ethylthioteleocidin B;

(xxvii) 14-deoxy-14-(2'-amino)ethylthioteleocidin B;

(xxviii) 14-deoxy-14-(3'-hydroxymethyl)phenylthioteleocidin B; and

(xxix) 14-deoxy-14-(2'-hydroxy)ethylthioteleocidin B.

EXAMPLE 3914-Deoxy-14-(N-methanesulfonyl)amino-7-octyl-(9S,12S)-indolactam V

To a solution of 4 mg of methanesulfonamide in 150 μL of methanolcontaining 1.3 mg of sodium methoxide was added approximately 6 mg of14-O-methanesulfonyl-7-octyl-(9S,12S)-indolactam V (prepared from(-)-7-octylindolactam V) in 300 μL of acetonitrile. After 26 h thismixture was diluted with ethyl acetate and washed twice with phosphatebuffers (pH 2 and pH 8). After drying over sodium sulfate, the organiclayer was concentrated under a nitrogen stream. Thin layerchromatographic analysis [silica; hexane/ethyl acetate (45:55)] showedthat the starting 14-O-methanesulfonyl-7-octyl-(9S,12S)-indolactam V(R_(f) =0.47) had been converted to14-deoxy-14-N-(methanesulfonyl)amino-7-octyl-(9S,12S)-indolactam V withR_(f) =0.38.

EXAMPLE 40 14-Deoxy-14-trimethylphosphonium-7-octyl-(9S,12S)-indolactamV, Methanesulfonate salt

To a solution of approximately 6 mg of14-O-methanesulfonyl-7-octyl-(9S,12S)-indolactam V (prepared from(-)-7-octylindolactam V) in 300 μL of acetonitrile was added 30 μL of 1Mtrimethylphosphine in toluene. After 26 h this mixture was concentratedunder a nitrogen stream. Thin layer chromatographic analysis [silica;methylene chloride/methanol (95:5)] showed that the staring14-O-methanesulfonyl-7-octyl-(9S,12S)-indolactam V (R_(f) =0.63) hadbeen converted to14-deoxy-14-trimethylphosphonium-7-octyl-(9S,12S)-indolactam V,methanesulfonate salt, with R_(f) =0.07.

EXAMPLE 41 14-Deoxy-14-triphenylphosphonium-7-octyl-(9S,12S)-indolactamV, Iodide salt

To a solution of approximately 5 mg of14-deoxy-14-iodo-7-octyl-(9S,12S)-indolactam V (prepared from14-O-methanesulfonyl-7-octyl-(9S,12S)-indolactam V) in 1 mL oftetrahydrofuran was added 6 mg of triphenyphosphine. After 2.5 h thismixture was concentrated under a nitrogen stream. Thin layerchromatographic analysis [silica; hexane/ethyl acetate (45:55)] showedthat the starting 14-deoxy-14-iodo-7-octyl-(9S,12S)-indolactam V (R_(f)=0.37) had been convened to14-deoxy-14-triphenylphosphonium-7-octyl-(9S,12S)-indolactam V, iodidesalt, with R_(f) =0.88.

EXAMPLE 42

In a similar manner the following compounds are prepared:

(i) 14-deoxy-14-tributylphosphonium-7-octylindolactam V, iodide salt;

(ii) 14-deoxy-14-triethylphosphonium-7-octylindolactam V, iodide salt;

(iii) 14-deoxy-14-methyldiphenylphosphonium-7-octylindolactam V, iodidesalt;

(iv) 14-deoxy-14-trimethylphosphonium-7-octyl-epi-indolactam V, iodidesalt;

(v) 14-deoxy-14-tributylphosphonium-7-octyl-epi-indolactam V, iodidesalt;

(vi) 14-deoxy-14-triethylphosphonium-7-octyl-epi-indolactam V, iodidesalt;

(vii) 14-deoxy-14-methyldiphenylphosphonium-7-octyl-epi-indolactam V,iodide salt;

(viii) 14-deoxy-14-trimethylphosphonium-6,7-tetramethyleneindolactam V,iodide salt;

(ix) 14-deoxy-14-tributylphosphonium-6,7-tetramethyleneindolactam V,iodide salt;

(x) 14-deoxy-14-triethylphosphonium-6,7-tetramethyleneindolactam V,iodide salt;

(xi) 14-deoxy-14-methyldiphenylphosphonium-6,7-tetramethyleneindolactamV, iodide salt;

(xii)14-deoxy-14-tributylphosphonium-7-octyl-12-des-iso-propyl-12-benzylindolactamV, iodide salt;

(xiii)14-deoxy-14-triethylphosphonium-7-octyl-12-des-iso-propyl-12-benzylindolactamV, iodide salt;

(xiv)14-deoxy-14-methyldiphenylphosphonium-7-octyl-12-des-iso-propyl-12-benzylindolactamV, iodide salt;

(xv)14-deoxy-14-trimethylphosphonium-7-octyl-12-des-iso-propyl-12-benzylindolactamV, iodide salt;

(xvi) 14-deoxy-14-tributylphosphoniumteleocidin B, iodide salt;

(xvii) 14-deoxy-14-triethylphosphoniumteleocidin B, iodide salt;

(xviii) 14-deoxy-14-methyldiphenylphosphoniumteleocidin B, iodide salt;and

(xix) 14-deoxy-14-trimethylphosphoniumteleocidin B, iodide salt.

EXAMPLE 43 14-Deoxy-14-trimethylsilyl-7-octyl-(9S,12S)-indolactam V

To a mixture of 10 mg of powdered zinc and 10 μL oftrimethylchlorosilane in 300 μL of anhydrous tetrahydrofuran was addedapproximately 5 mg of 14-deoxy-14-iodo-7-octyl-(9S,12S)-indolactam V(prepared from 14-O-methanesulfonyl-7-octyl-(9S,12S)-indolactam V) in500 μL of tetrahydrofuran. After 2.5 h, thin layer chromatographicanalysis [silica; hexane/ethyl acetate (45:55)] showed that the starting14-deoxy-14-iodo-7-octyl-(9S,12S)-indolactam V (R_(f) =0.37) had beenlargely converted to14-deoxy-14-trimethylsilyl-7-octyl-(9S,12S)-indolactam V with R_(f)=0.49.

EXAMPLE 44

In a similar manner the following compounds are prepared:

(i) 14-deoxy-14-trimethylsilyl-7-octyl-epi-indolactam V;

(ii)14-deoxy-14-trimethylsilyl-7-octyl-12-des-iso-propyl-12-benzylindolactamV;

(iii) 14-deoxy-14-trimethylsilyl-6,7-tetramethyleneindolactam V; and

(iv) 14-deoxy-14-trimethylsilylteleocidin B.

EXAMPLE 45 20-Deoxy-20-hydroximinophorbol12,13-Bis[(2',4'-difluorophenyl)acetate] and20-Deoxy-3-deoxo-3,20-bis(hydroximino)phorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 250 mg of 20-deoxy-20-oxophorbol12,13-bis[(2',4'-difluorophenyl)acetate] [G. Kreibich and E. Hecker, Z.Krebsforsch., 74, 448-456 (1970)] in 6 mL of methanol was added 4 mL ofa solution prepared by adding 40 mL of 0.375M sodium methoxide inmethanol to 2.5 g of hydroxylamine hydrochloride. After 2.5 h themixture was concentrated in vacuo and the residue was partitionedbetween water and ethyl acetate. The organic layer was filtered througha funnel containing layers of sodium chloride, sodium sulfate and silicagel (N/N/S) and concentrated in vacuo. After purification of the residueby preparative liquid chromatography [silica; methylene chloride/ethylacetate (85:15)], 211 mg of 20-deoxy-20-hydroximinophorbol12,13-bis[(2',4'-difluorophenyl)acetate] and 27 mg of20-deoxy-3-deoxo-3,20-bis(hydroximino)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] were obtained. The structureswere confirmed by NMR and mass spectral analysis.

EXAMPLE 46

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-hydroximinophorbol 12,13-didecanoate;

(ii) 20-deoxy-20-hydroximinophorbol 12,13-dibutyrate;

(iii) 12,20-dideoxy-20-hydroximinophorbol 13-decanoate;

(iv) 12,20-dideoxy-20-hydroximinophorbol13-(2',4'-difluorophenyl)acetate;

(v) 20-deoxy-20-hydroximinophorbol 12-myristate 13-acetate; and

(vi) 20-deoxy-3-deoxo-3,20-bis(hydroximino)phorbol 12-myristate13-acetate.

EXAMPLE 47 20-Deoxy-20-methoximinophorbol12,13-Bis[(2',4'-difluorophenyl)acetate] and20-Deoxy-3-deoxo-3,20-bis(methoximino)phorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 100 mg of 20-deoxy-20-oxophorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 1 mL of pyridine was added35 mg of methoxylamine hydrochloride. After 2.5 h the mixture waspartitioned between ethyl acetate and phosphate buffer (pH 2). Theorganic layer was washed with phosphate buffer (pH 8), filtered throughN/N/S and concentrated in vacuo. After preparative liquid chromatography[silica; hexane/ethyl acetate (80:20)] of the residue, 96 mg of20-deoxy-20-methoximinophorbol 12,13-bis[(2',4'-difluorophenyl)acetate]and 4 mg of 20-deoxy-3-deoxo-3,20-bis(methoximino)phorbol 12,13-bis[(2',4'-difluorophenyl)acetate] were obtained. The structures wereconfirmed by NMR and high resolution mass spectral analysis.

EXAMPLE 48

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-hydroximinophorbol 12-(3'-phenoxy)benzoate 13-butyrate;

(ii) 20-deoxy-20-hydroximinophorbol 12-(3',5'-difluorophenyl)acetate13-(3',4'-difluorobenzoate);

(iii) 20-deoxy-20-hydroximinophorbol 12-(3',5'-difluorocinnamate)13-[3'-(2",4"-difluorophenyl)propionate];

(iv) 20-deoxy-20-hydroximinophorbol12,13-bis[(2',4'-dichlorophenyl)acetate];

(v) 20-deoxy-20-methoximinophorbol 12,13-didecanoate;

(vi) 12,20-dideoxy-20-methoximinophorbol 13-decanoate;

(vii) 20-deoxy-20-t-butoximinophorbol 12-myristate 13-acetate;

(viii) 20-deoxy-20-carboxymethoximinophorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(ix) 20-deoxy-20-(4-nitrobenzoximino)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(x) 20-deoxy-20-alloximinophorbol 12-myristate 13-acetate; and

(xi) 20-deoxy-20-oxophorbol12,13-bis[(2',4'-difluorophenyl)acetate]20-semicarbazone.

EXAMPLE 49 6-Deshydroxymethyl-6-cyanophorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

A solution of 100 mg of 20-deoxy-20-hydroximinophorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 0.5 mL pyridine was added toa mixture of 9 mg cupric sulfate and 35 mg of triethylamine in 1.5 mL ofmethylene chloride. After 1 h 47 mg of dicyclohexylcarbodiimide wasadded to the green solution. After 3 h at room temperature the reactionmixture was heated at 40°-45° C. for 3 h. During this period another 50mg of dicyclohexylcarbodiimide was added. After cooling the mixture toroom temperature, 0.5 mL of formic acid was added followed bypartitioning between ethyl acetate and phosphate buffer (pH 2). Theorganic layer was washed with phosphate buffer (pH 8), filtered throughN/N/S and concentrated in vacuo. Preparative liquid chromatography[silica; hexane/ethyl acetate (75:25)] afforded 43 mg of6-deshydroxymethyl-6-cyanophorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and high resolution mass spectral analysis.

EXAMPLE 50

In a similar manner the following compounds are prepared:

(i) 6-deshydroxymethyl-6-cyanophorbol 12-(3',5'-difluorophenyl)acetate13-(3',4'-difluorobenzoate);

(ii) 6-deshydroxymethyl-6-cyanophorbol 12,13-didecanoate; and

(iii) 12-deoxy-6-deshydroxymethyl-6-cyanophorbol 13-decanoate.

EXAMPLE 51 6-Deshydroxymethyl-6-carboxyphorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 1.51 g of 20-deoxy-2-oxo-phorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 55 mL of methylene chloride,150 mL of t-butyl alcohol and 10 mL of 2-methyl-2-butene was added 18.75mL of a solution prepared by dissolving 1.99 g of sodium chlorite and 2g of potassium dihydrogen phosphate in 20 mL of deionized water. After1.25 h about 10 mL of 20% aq. sodium thiosulfate was added and themixture was partially concentrated in vacuo. The residue was partitionedbetween ethyl acetate and phosphate buffer (pH 8). The organic layer waswashed sequentially with pH 2 and pH 8 phosphate buffers and dried oversodium sulfate. After concentration in vacuo, 1.53 g of6-deshydroxymethyl-6-carboxyphorbol12,13-bis[(2',4'-difluorophenyl)acetate] was obtained. An analyticalsample was obtained by preparative liquid chromatography [silica;methylene chloride/methanol (95:5)] and the structure confirmed by NMRand mass spectral analysis.

EXAMPLE 52

In a similar manner the following compounds are prepared:

(i) 6-deshydroxymethyl-6-carboxyphorbol 12,13-dibutyrate;

(ii) 6-deshydroxymethyl-6-carboxyphorbol 12,13-didecanoate;

(iii) 6-deshydroxymethyl-6-carboxy-3-deoxo-3-benzoyloxyphorbol12-butyrate 13-(2',4'-difluorobenzoate);

(iv)6-deshydroxymethyl-6-carboxy-3-deoxo-3-(2',4'-difluorophenylacetoxy)phorbol12-(2',4'-difluorophenyl)acetate 13-(4'-biphenyl)acetate;

(v) 12-deoxy-6-deshydroxymethyl-6-carboxyphorbol 13-decanoate;

(vi) 12-deoxy-6-deshydroxymethyl-6-carboxyphorbol13-(2',4'-difluorophenyl)acetate;

(vii) 6-deshydroxymethyl-6-carboxyphorbol 12-myristate 13-acetate;

(viii) 6-deshydroxymethyl-6-carboxyphorbol 12-(3'-phenoxy)benzoate13-butyrate;

(ix) 6-deshydroxymethyl-6-carboxyphorbol12-(3',5'-difluorophenyl)acetate 13-(3',4'-difluorobenzoate);

(x) 6-deshydroxymethyl-6-carboxyphorbol 12-(3',5'-difluorocinnamate)13-[3'-(2",4"-difluorophenyl)propionate];

(xi) 6-deshydroxymethyl-6-carboxyphorbol12,13-bis[(2',4'-dichlorophenyl)acetate];

(xii) 6-deshydroxymethyl-6-carboxyresiniferonol9,13,14-orthophenylacetate;

(xiii) 5-O-trimethylsilyl-6-deshydroxymethyl-6-carboxyingenol3,4-acetonide;

(xiv) 6-deshydroxymethyl-6-carboxyphorbol 12-octyldimethylsilylacetate13-(2',4'-difluorophenyl)acetate; and

(xv) 6-deshydroxymethyl-6-carboxy-12-deoxyphorbol13-(2',4'-difluorophenyl)acetate.

EXAMPLE 53 6-Deshydroxymethyl-6-methoxycarbonylphorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of approximately 100 mg of6-deshydroxymethyl-6-carboxyphorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 10 mL of tetrahydrofuran wasadded an excess of diazomethane in ether. After about one hour thesolution was concentrated in vacuo and the residue subjected topreparative liquid chromatography [silica; hexane/ethyl acetate (70:30)]to afford 96 mg of 6-deshydroxymethyl-6-methoxycarbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and mass spectral analysis.

EXAMPLE 54

In a similar manner the following compounds are prepared:

(i) 6-deshydroxymethyl-6-methoxycarbonylphorbol 12,13-didecanoate; and

(ii) 12-deoxy-6-deshydroxymethyl-6-methoxycarbonylphorbol 13-decanoate.

EXAMPLE 55 6-Deshydroxymethyl-6-(N-succinimidyloxy)carbonylphorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

A solution of 580 mg of dicyclohexylcarbodiimide in 20 mL oftetrahydrofuran was added to an ice cooled mixture of 256 mg ofN-hydroxysuccinimide and 1.38 g of 6-deshydroxymethyl-6-carboxyphorbol12,13-bis[(2',4'-difluorophenyl)acetate] in acetonitrile (85mL)/tetrahydrofuran (50 mL). After 10 min the mixture was allowed towarm to room temperature under positive nitrogen pressure. After 7 h themixture was filtered and then concentrated in vacuo. Preparative liquidchromatography [silica; hexane/ethyl acetate (50:50)] afforded 1.53 g of6-deshydroxymethyl-6-(N-succinimidyloxy)carbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate], the structure of which wasconfirmed by NMR spectral analysis.

EXAMPLE 56

In a similar manner the following compounds are prepared:

(i) 12-deoxy-6-deshydroxymethyl-6-(N-succinimidyloxy)carbonylphorbol13-decanoate;

(ii) 6-deshydroxymethyl-6-(N-succinimidyloxy)carbonylphorbol12-myristate 13-acetate; and

(iii) 6-deshydroxymethyl-6-(N-succinimidyloxy)carbonylphorbol12-(3'-phenoxy)benzoate 13-butyrate.

EXAMPLE 576-Deshydroxymethyl-6-[N-(2',3'-dihydroxypropyl)carboxamido]phorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

Over 2.5 h a total of about 50 mg of ()-3-amino-1,2-propandiol was addedto a solution of 100 mg of6-deshydroxymethyl-6-(N-succinimidyloxy)carbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate] in tetrahydrofuran (1mL)/acetonitrile (0.5 mL). After another 0.75 h the mixture waspartitioned between phosphate buffer (pH 2) and ethyl acetate. Theorganic layer was washed with phosphate buffer (pH 8), dried over sodiumsulfate and concentrated in vacuo. The resulting residue was subjectedto preparative liquid chromatography [silica; methylenechloride/methanol (94:6) to afford 78 mg of6-deshydroxymethyl-6-[N-(2',3'-dihydroxypropyl)carboxamido]phorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and mass spectral analysis.

EXAMPLE 58

In a similar manner the following compounds are prepared:

(i)12-deoxy-6-deshydroxymethyl-6-[N-(2'-hydroxyethyl)carboxamido]phorbol13-decanoate;

(ii)6-deshydroxymethyl-6-[N-(2'S)-1'-hydroxyprop-2'-yl)carboxamido]phorbol12-myristate 13-acetate;

(iii)6-deshydroxymethyl-6-(2'-hydroxymethylpiperidin-1'-yl)carbonylphorbol12-(3'-phenoxy)benzoate 13-butyrate; and

(iv) 6-deshydroxymethyl-6-carboxamidophorbol12,13-bis[(2',4'-difluorophenyl)acetate].

EXAMPLE 59 6-Deshydroxymethyl-6-(2'-hydroxy)ethoxycarbonylphorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

A solution consisting of 100 mg of6-deshydroxymethyl-6-(N-succinimidyloxy)carbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate], a few mg of4-dimethylaminopyridine and a large excess of ethylene glycol in 1 mL oftetrahydrofuran was allowed to stand for 6 days at room temperature. Thesolution was then partitioned between water and ethyl acetate, theorganic layer dried over sodium sulfate and concentrated in vacuo. Afterpurification by preparative liquid chromatography [silica; hexane/ethylacetate], 32 mg of6-deshydroxymethyl-6-(2'-hydroxy)ethoxycarbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate] was obtained. The structure wasconfirmed by NMR and mass spectral analysis.

EXAMPLE 60

In a similar manner the following compounds are prepared:

(i) 12-deoxy-6-deshydroxymethyl-6-(2'-hydroxy)ethoxycarbonylphorbol13-decanoate; and

(ii) 6-deshydroxymethyl-6-(3'-hydroxy)propyloxycarbonylphorbol12-myristate 13-acetate.

EXAMPLE 61 20-Deoxy-20-(3'-hydroxypropylthio)phorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 100 mg of 20-deoxy-20-chlorophorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 1 mL of acetonitrile wasadded a solution of 37 μL of 3-mercaptopropanol in 383 μL of 0.43Mmethanolic sodium methoxide. After about 15 min the reaction mixture waspartitioned between ethyl acetate and phosphate buffer (pH 8). Theorganic layer was dried over sodium sulfate and concentrated under astream of nitrogen. The residue was chromatographed silica; hexane/ethylacetate (65:35)] to afford 77 mg of20-deoxy-20-(3'-hydroxypropylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate].

EXAMPLE 62

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(ii) 20-deoxy-20-(2',3'-dihydroxypropylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(iii) 20-deoxy-20-(4'-hydroxy-n-butylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(iv) 20-deoxy-20-(2'-hydroxypropylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(v) 20-deoxy-20-(2'-aminoethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(vi) 20-deoxy-20-[2'-(formylamino)ethylthio]phorbol 12-myristate13-acetate;

(vii) 20-deoxy-20-[2'-(acetylamino)ethylthio]phorbol 12-myristate13-acetate;

(viii) 20-deoxy-20-[2'-(methylsulfonylamino)ethylthio]phorbol12-myristate 13-acetate;

(ix) 20-deoxy-20-propylthiophorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(x) 20-deoxy-20-(2'-mercaptoethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(xi) 20-deoxy-20-[2'-(hydroxymethyl)phenylthio]phorbol 12-myristate13-acetate;

(xii) 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12,13-dibenzoate;

(xiii) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis(phenylacetate);

(xiv) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(pentafluorophenyl)acetate];

(xv) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(2',4'-dichlorophenyl)acetate];

(xvi) 20-deoxy-20-(2',3'-dihydroxypropylthio)phorbol12-4'-(9",10"-dihydrophenanthrene-2")butyrate]13-(2',4'-difluorobenzoate);

(xvii) 20-deoxy-20-(3'-hydroxypropylthio)phorbol12-(2',4'-difluorocinnamate) 13-(2',4'-difluorophenyl)acetate;

(xviii) 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-(4'-biphenylacetate)13-[3'-(3",5"-difluorophenyl)propionate];

(xix) 20-deoxy-20-(2'-carboxamidoethylthio)phorbol12,13-bis[(2',4'-dichlorophenyl)acetate];

(xx) 20-deoxy-20-(2'-hydroxyethylthio)-12-O-,13-O-bis(isopropyldimethylsilyl)phorbol;

(xxi) 20-deoxy-20-(4'-hydroxy-2'-butenylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(xxii) 12,20-dideoxy-20-(2'-hydroxyethylthio)phorbol13-(2',4'-difluorophenyl)acetate;

(xxiii) 20-deoxy-20-(2'-hydroxyethylthio)resiniferonol9,13,14-orthophenylacetate;

(xxiv) 20-deoxy-20-phenylselenophorbol12,13-bis[(2',4'-difluorophenyl)acetate]; and

(xxv) 20-deoxy-20-methylselenophorbol12,13-bis[(2',4'-difluorophenyl)acetate] (sodium methylselenide isprepared by treatment of dimethyldiselenide with sodium borohydride).

EXAMPLE 63 20-Deoxy-20-[2'-(t-butyldiphenylsilyloxy)ethylthio]phorbol

A solution containing 12 g of 2-hydroxyethyldisulfide and 12.2 g ofimidazole in 150 mL of dried N,N-dimethylformamide was treated with41.65 g of t-butyldiphenylchlorosilane. After 2 h, another 25 g of thesilane and 5 g of imidazole were added along with 10 mL of pyridine.After another 3.5 h the mixture was partitioned between ethyl acetateand water. The organic layer was then washed with phosphate buffer (pH 2and pH 8, sequentially), filtered through N/N/S and concentrated invacuo. Preparative liquid chromatography [silica; hexane/methylenechloride (90:10)] afforded about 46 g of2-t-butyldiphenylsilyloxyethyldisulfide.

To a solution of 46 g of 2-t-butyldiphenylsilyloxyethyldisulfide in 650mL of tetrahydrofuran was added a mixture of 40 g of zinc dust and 10 mLof acetic acid in 50 mL of tetrahydrofuran. After 2 h the supernatantwas decanted into 2 L of water and the residue washed twice with 500 mLof ethyl acetate each. The aqueous solution was extracted with ethylacetate and the combined organic layers washed with brine, filteredthrough N/N/S and concentrated in vacuo. The residue was dissolved inhexane/methylene chloride (85:15) and filtered through silica.Concentration in vacuo afforded 37.5 g of2-t-butyldiphenylsilyloxyethylthiol.

A solution prepared from 1 g of sodium and 17 g of2-t-butyldiphenylsilyloxyethylthiol in 100 mL of methanol was added toapproximately 4 g of crude 20-deoxy-20-chlorophorbol in 500 mL of ethylacetate. After 35 min, a dilute phosphate buffer (pH 2) was added andthen the organic layer was washed with phosphate buffer (pH 8), filteredthrough N/N/S and concentrated in vacuo. Preparative liquidchromatography [silica; hexane/ethyl acetate (25:75)] of the residueafforded 6.0 g of20-deoxy-20-[2'-(t-butyldiphenylsilyloxy)ethylthio]phorbol.

EXAMPLE 64 20-Deoxy-20-(2'-hydroxyethylthio)phorbol12,13-Bis[3-(pentafluorophenyl)propionate]

To an ice-cold solution of 300 mg of 1,1'-carbonyldiimidazole in 3 mLnitromethane was added 420 μL of methyl triflate followed by 441 mg of3-(pentafluorophenyl)propionic acid in 5 mL of nitromethane. After 5 mina solution of 245 mg of20-deoxy-20-[2'-(t-butyldiphenylsilyloxy)ethylthio]phorbol and 30 mg of4-dimethylaminopyridine in 1.5 mL of anhydrous tetrahydrofuran wasadded. This mixture was allowed to stand at room temperature for 17 hthen it was partitioned between ethyl acetate and phosphate buffer (pH8). After drying (over sodium sulfate) the organic layer wasconcentrated in vacuo. By analysis of the thin layer chromatogram thisresidue was found to consist of a mixture of the mono and diesters. Theresidue was again subjected to the above conditions for 2 h and thentreated as before. Preparative liquid chromatography [silica;hexane/ethyl acetate (85:15)] afforded 320 mg of20-deoxy-20-[2'-(t-butyldiphenylsilyloxy)ethylthio]phorbol12,13-bis[3'-(pentafluorophenyl)propionate].

Three hundred μL of 1M tetrabutylammonium fluoride in tetrahydrofuranwas added to a solution of 250 mg of20-deoxy-20-[2'-(t-butyldiphenylsilyloxy)ethylthio]phorbol12,13-bis[3'-(pentafluorophenyl)propionate] in 4 mL of tetrahydrofuran.After 2 h the solution was partitioned between phosphate buffer (pH 8)and ethyl acetate. The organic layer was filtered through N/N/S andconcentrated in vacuo. Preparative liquid chromatography [silica;hexane/ethyl acetate (60:40)] afforded 66 mg of20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[3'-(pentafluorophenyl)propionate]. This structure wasconfirmed by NMR and mass spectral analysis.

EXAMPLE 65

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis(2',4'-difluorobenzoate);

(ii) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(3',4'-dimethoxyphenyl)acetate];

(iii) 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12,13-didecanoate(decanoyl anhydride is used as the acylating agent in the presence ofpyridine and 4-dimethylaminopyridine);

(iv) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis(diphenylphosphate) (diphenylchlorophosphate is used as theacylating agent in the presence of triethylamine and4-dimethylaminopyridine);

(v) 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12,13-bis(diethylphosphate)(diethylchlorophosphate is used as the acylating agent in the presenceof triethylamine and 4-dimethylaminopyridine); and

(vi) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)carbamate] (2,4-difluorophenylisocyanate is used as the carbamoylating agent in the presence ofdibutyltin dilaurate and 4-dimethylaminopyridine).

EXAMPLE 66 20-Deoxy-20-(2'-carboxyethylthio)phorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

Fifty μL of 3-mercaptopropionic acid was added to 1.6 mL of 0.435Mmethanolic sodium methoxide. This solution was then added to 100 mg of20-deoxy-20-chlorophorbol 12,13-bis[(2',4'-difluorophenyl)acetate] in 2mL of dried acetonitrile. After 0.5 h the mixture was partitionedbetween ethyl acetate and phosphate buffer (pH 2). The organic layer wasthen washed with phosphate buffer (pH 8), filtered through N/N/S andconcentrated in vacuo. Purification by preparative liquid chromatography[silica; methylene chloride/methanol (97:3)] afforded 54 mg of20-deoxy-20-(2'-carboxyethylthio)phorbol12,13-bis(2',4'-difluorophenylacetate). This structure was confirmed byNMR and mass spectral analysis.

EXAMPLE 67

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-(2'-carboxyethylthio)phorbol 12-(4'-biphenyl)acetate13-[3'-(3", 5"-difluorophenyl)propionate];

(ii) 20-deoxy-20-(2'-carboxyethylthio)phorbol 12-myristate 13-acetate;

(iii) 12,20-deoxy-20-(2'-carboxyethylthio)phorbol13-(2',4'-difluorophenyl)acetate;

(iv) 20-deoxy-20-(2'-carboxyethylthio)phorbol 12-(4'-biphenyl)acetate13-[3'-(3",5"-difluorophenyl)propionate];

(v) 20-deoxy-20-(2'-carboxyethylthio)phorbol 12-myristate 13-acetate;

(vi) 12,20-dideoxy-20-(2'-carboxyethylthio)phorbol13-(2',4'-difluorophenyl)acetate;

(vii) 20-deoxy-20-(2'-carboxyethylthio)resiniferonol9,13,14-orthophenylacetate; and

(viii) 20-deoxy-20-[(3'-hydroxylamino-3'-oxopropyl)thio]phorbol12-myristate 13-acetate.

EXAMPLE 68 20-Deoxy-20-mercaptophorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

A solution of 72 mg of hydrated sodium hydrogensulfide in 1 mL ofmethanol and 2 μL of acetic acid was prepared. Two hundred and fifty μLof this solution was added to a solution of 100 mg of20-deoxy-20-chlorophorbol 12,13-bis[(2',4'-difluorophenyl)acetate] in 1mL of acetonitrile. After 35 min another 100 μL of the solution wasadded. After 25 min phosphate buffer (pH 6) was added and the mixturewas extracted with ethyl acetate. The organic layer was filtered throughN/N/S and concentrated in vacuo. The residue was subjected topreparative liquid chromatography [silica; hexane/ethyl acetatemixtures] to afford 32 mg of 20-deoxy-20-mercaptophorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and mass spectral analysis.

EXAMPLE 69

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-mercaptophorbol 12-(4'-biphenyl)acetate13-[3'-(3",5"-difluorophenyl)propionate];

(ii) 20-deoxy-20-mercaptophorbol 12-myristate 13-acetate;

(iii) 12,20-dideoxy-20-mercaptophorbol 13-(2',4'-difluorophenyl)acetate;and

(iv) 20-deoxy-20-mercaptoresiniferonol 9,13,14-orthophenylacetate.

EXAMPLE 70 20-Deoxy-20-acetonylthiophorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a mixture of chloroacetone and potassium iodide in acetonitrile isadded 20-deoxy-20-mercaptophorbol12,13-bis[(2',4'-difluorophenyl)acetate]. After a period of time themixture is partitioned between ethyl acetate and phosphate buffer (pH 8)and the organic layer is filtered through N/N/S and concentrated invacuo. Purification by liquid chromatography affords20-deoxy-20-acetonylthiophorbol12,13-bis[(2',4'-difluorophenyl)acetate].

EXAMPLE 71 20-Deoxy-20-(2'-hydroxyethylsulfinyl)phorbol 12-Myristate13-Acetate and 20-Deoxy-20-(2'-hydroxyethylsulfonyl)phorbol 12-Myristate13-Acetate

To a solution of 100 mg of 20-deoxy-20-(2'-hydroxyethylthio)phorbol12-myristate 13-acetate in 2 mL of t-butyl alcohol was added 100 μL of30% hydrogen peroxide. After 30 min, 2 mL of 20% aq. sodium thiosulfatewas added and the mixture partitioned between water and ethyl acetate.The organic layer was dried over sodium sulfate and concentrated invacuo. After preparative liquid chromatography [silica; methylenechloride/isopropyl alcohol] on the residue, 67 mg of20-deoxy-20-(2'-hydroxyethylsulfinyl)phorbol 12-myristate 13-acetate and14 mg of 20-deoxy-20-(2'-hydroxyethylsulfonyl)phorbol 12-myristate13-acetate were obtained. The structures were confirmed by NMR and massspectral analysis.

EXAMPLE 72

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-(2'-hydroxyethylsulfinyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(ii) 20-deoxy-20-(2'-hydroxyethylsulfinyl)phorbol12,13-bis[(pentafluorophenyl)acetate];

(iii) 20-deoxy-20-(propylsulfinyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(iv) 20-deoxy-20-(2'-hydroxyethylsulfinyl)phorbol12,13-bis[3'-(pentafluorophenyl)propionate];

(v) 20-deoxy-20-(2'-hydroxyethylsulfinyl)-3-deoxo-3-hyroximinophorbol12,13-bis[3'-(pentafluorophenyl)propionate];

(vi) 20-deoxy-20-(2'-carboxyethylsulfinyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; and

(vii) 20-deoxy-20-(3'-hydroxypropylsulfinyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate].

EXAMPLE 73 20-Deoxy-20-(2'-hydroxyethylsulfonyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]

To a solution of 100 mg of 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 1 mL of methylene chloridewas added, drop wise, 49 mg of m-chloroperbenzoic acid in 1 mL ofmethylene chloride. After about one hour another 28 mg of peracid wasadded. A few minutes later the reaction mixture was partitioned betweenaq. sodium thiosulfate (approximately 5%) and ethyl acetate. The organiclayer was dried over sodium sulfate and concentrated in vacuo.Preparative liquid chromatography [silica; hexane/ethyl acetate (40:60)]of the residue afforded 69 mg of20-deoxy-20-(2'-hydroxyethylsulfonyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and mass spectral analysis.

EXAMPLE 74

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-(2'-hydroxyethylsulfonyl)phorbol12,13-bis[(pentafluorophenyl)acetate];

(ii) 20-deoxy-20-(propylsulfonyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(iii) 20-deoxy-20-(2'-hydroxyethylsulfonyl)phorbol12,13-bis[3'-(pentafluorophenyl)propionate];

(iv) 20-deoxy-20-(2'-hydroxyethylsulfonyl)-3-deoxo-3-hyroximinophorbol12,13-bis[3'-(pentafluorophenyl)propionate];

(v) 20-deoxy-20-(2'-carboxyethylsulfonyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(vi) 20-deoxy-20-(3'-hydroxypropylsulfonyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; and

(vii) 20-deoxy-20-(2'-hydroxyethylsulfonyl)phorbol 12-myristate13-acetate.

EXAMPLE 75 20-Deoxy-20-cyanophorbol 12-Myristate 13-Acetate

A mixture consisting of 500 mg of 20-deoxy-20-chlorophorbol 12-myristate13-acetate, 80 mg of potassium cyanide, 150 mg of tetrabutylammoniumchloride, 75 mg of 18-crown-6,3 mL of chloroform and 2.8 mL of water wasstirred for 22 h at room temperature. The mixture was then partitionedbetween ethyl acetate and phosphate buffer (pH 8) and the organic layerwas filtered through N/N/S and concentrated in vacuo. Preparative liquidchromatography [silica; hexane/isopropyl alcohol (92:8)] afforded20-deoxy-20-cyanophorbol 12-myristate 13-acetate.

EXAMPLE 76

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-cyanophorbol 12,13-bis(phenylacetate);

(ii) 20-deoxy-20-cyanophorbol 12,13-bis[(pentafluorophenyl)acetate];

(iii) 20-deoxy-20-cyanophorbol 12,13-bis[(2',4'-dichlorophenyl)acetate];

(iv) 20-deoxy-20-cyanophorbol12-4'-(9",10"-dihydrophenanthrene-2")butyrate]13-(2',4'-difluorobenzoate);

(v) 20-deoxy-20-cyanophorbol 12-(2',4'-difluorocinnamate)13-(2',4'-difluorophenylacetate);

(vi) 20-deoxy-20-cyanophorbol 12-(4'-biphenyl)acetate13-[3'-(3",5"-difluorophenyl)propionate];

(vii) 12,20-dideoxy-20-cyanophorbol 13-(2',4'-difluorophenyl)acetate;and

(viii) 20-deoxy-20-cyanoresiniferonol 9,13,14-orthophenylacetate.

EXAMPLE 77 20-Deoxy-20-azidophorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 100 mg of 20-deoxy-20-chlorophorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 1 mL of anhydrous methanolwas added 88 mg of sodium azide. After 3 h the mixture was partitionedbetween water and ethyl acetate. The organic layer was filtered throughN/N/S and concentrated in vacuo. Upon further purification bychromatography [silica; hexane/ethyl acetate (80:20)], 88 mg of20-deoxy-20-azidophorbol 12,13-bis[(2',4'-difluorophenyl)acetate] wasobtained. The structure was confirmed by NMR and mass spectral analysis.

EXAMPLE 78

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-azidophorbol 12,13-bis(phenylacetate);

(ii) 20-deoxy-20-azidophorbol 12,13-bis[(pentafluorophenyl)acetate];

(iii) 20-deoxy-20-azidophorbol 12,13-bis[(2',4'-dichlorophenyl)acetate];

(iv) 20-deoxy-20-azidophorbol 12-4'-(9",10"-dihydrophenanthrene-2")butyrate]13-(2',4'-difluorobenzoate);

(v) 20-deoxy-20-azidophorbol 12-(2',4'-difluorocinnamate)13-(2',4'-difluorophenylacetate);

(vi) 20-deoxy-20-azidophorbol 12-(4'-biphenyl)acetate13-[3'-(3",5"-difluorophenyl)propionate];

(vii) 12,20-dideoxy-20-azidophorbol 13-(2',4'-difluorophenyl)acetate;

(viii) 12,20-dideoxy-20-azidophorbol 13-decanoate;

(ix) 20-deoxy-20-azidoresiniferonol 9,13,14-orthophenylacetate;

(x) 20-deoxy-20-azidophorbol 12-myristate 13-acetate;

(xi) 20-deoxy-20-azidophorbol 12,13-dibenzoate;

(xii) 20-deoxy-20-azido-12-O-,13-O-bis(isopropyldimethylsilyl)phorbol;

(xiii) 20-deoxy-20-azidophorbol 12,13-bis(2',4'-difluorobenzoate);

(xiv) 20-deoxy-20-azidophorbol12,13-bis[(3',4'-dimethoxyphenyl)acetate];

(xv) 20-deoxy-20-azidophorbol 12,13-didecanoate;

(xvi) 5-O-trimethylsilyl-20-deoxy-20-azidoingenol 3,4-acetonide;

(xvii) 20-deoxy-20-azidophorbol 12-octyldimethylsilylacetate13-(2',4'-difluorophenylacetate); and

(xviii) 20-deoxy-20-azidophorbol 12-(2',4'-difluorophenylacetate)13-diphenylphosphate.

EXAMPLE 79 20-Deoxy-20-aminophorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

A solution of 47.5 mg of20-deoxy-20-azidophorbol12,13-bis[(2',4'-difluorophenyl)acetate]and 27 mg of triphenylphosphinein 1 mL of tetrahydrofuran was stirred for 4.5 h at room temperature. Atthis time 10 mg of triphenylphosphine was added followed 1.5 h later by100 μL of water and 1.5 h after that with a few mg of silica. Afteranother 1.5 h the mixture was filtered through silica gel andconcentrated by a stream of nitrogen. Preparative liquid chromatographyafforded 18 mg of 20-deoxy-20-aminophorbol12,13-bis[(2',4'-difluorophenyl)acetate].

EXAMPLE 80

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-aminophorbol 12,13-bis(phenylacetate);

(ii) 20-deoxy-20-aminophorbol 12,13-bis[(pentafluorophenyl)acetate];

(iii) 20-deoxy-20-aminophorbol 12,13-bis[(2',4'-dichlorophenyl)acetate];

(iv) 20-deoxy-20-aminophorbol12-4'-(9",10"-dihydrophenanthrene-2")butyrate]13-(2',4'-difluorobenzoate);

(v) 20-deoxy-20-aminophorbol 12-(4'-biphenyl)acetate13-[3'-(3",5"-difluorophenyl)propionate];

(vi) 12,20-dideoxy-20-aminophorbol 13-(2',4'-difluorophenyl)acetate;

(vii) 20-deoxy-20-aminoresiniferonol 9,13,14-orthophenylacetate;

(viii) 20-deoxy-20-aminophorbol 12-myristate 13-acetate;

(ix) 20-deoxy-20-aminophorbol 12,13-dibenzoate;

(x) 20-deoxy-20-aminophorbol 12,13-bis(2',4'-difluorobenzoate); and

(xi) 20-deoxy-20-aminophorbol 12,13-bis[(3',4'-dimethoxyphenyl)acetate].

EXAMPLE 81 20-Deoxy-20-(N'-methylureido)phorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 50 mg of 20-deoxy-20-aminophorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 1 mL of tetrahydrofuran isadded 20 μL of methyl isocyanate. After several hours some methanol isadded to the reaction mixture to consume excess isocyanate and themixture is concentrated. Purification by liquid chromatography affords20-deoxy-20-(N'-ureido)phorbol 12,13-bis[(2',4'-difluorophenyl)acetate].

EXAMPLE 82

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-(N'-ethylureido)phorbol 12-myristate 13-acetate;

(ii) 20-deoxy-20-(N'-propylureido)phorbol 12,13-dibenzoate;

(iii) 20-deoxy-20-(N'-methylthioureido)phorbol12,13-bis(2',4'-difluorobenzoate);

(iv) 12,20-dideoxy-20-(N'-butylureido)phorbol13-(2',4'-difluorophenyl)acetate;

(v) 20-deoxy-20-[N'-(3'-methoxyphenyl)ureido]phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(vi) 20-deoxy-20-[N'-(3'-trifluoromethylphenyl)ureido]phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(vii) 20-deoxy-20-(N'-allylthioureido)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(viii) 20-deoxy-20-(N'-phenethylthioureido)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; and

(ix) 20-deoxy-20-(N'-methylureido)resiniferonol9,13,14-orthophenylacetate.

EXAMPLE 83 20-Deoxy-20-bromophorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 1.0 g of phorbol12,13-bis[(2',4'-difluorophenyl)acetate] and 1.58 g of 2,6-lutidine in20 mL of dry acetonitrile was added 3.21 g ofdibromotriphenylphosphorane. After 0.5 h approximately 10 mL of waterwas added and the mixture was extracted with ethyl acetate. The combinedorganic layers were washed with phosphate buffers, pH 2 and pH 8sequentially, and then brine before being filtered through N/N/S andconcentrated in vacuo.

Preparative liquid chromatography [silica; hexane/ethyl acetate (75:25)]afforded 665 mg of 20-deoxy-20-bromophorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and mass spectral analysis.

EXAMPLE 84

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-bromophorbol 12-myristate 13-acetate;

(ii) 20-deoxy-20-bromophorbol 12-(4'-biphenyl)acetate 13-acetate; and

(iii) 20-deoxy-20-bromophorbol12-[(2',2'-dimethyl)(2",4"-difluorophenyl)acetate 13-butyrate.

EXAMPLE 85 20-Deoxy-20-[N,N-bis(2'-hydroxyethyl)amino]phorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 100 mg of 20-deoxy-20-bromophorbol12,13-bis[(2',4'-difluorophenyl)acetate] and 11 μL of pyridine in 1 mLof acetonitrile was added, over a 5 h period, 50 mg of diethanolamine in1 mL of acetonitrile. The reaction was then partitioned between ethylacetate and phosphate buffer (pH 2). The organic layer was washed withphosphate buffer (pH 8), dried over sodium sulfate and concentrated invacuo. After preparative liquid chromatography [silica; methylenechloride/tetrahydrofuran (75:25)], 51 mg of20-deoxy-20-[N,N-bis(2'-hydroxyethyl)amino]phorbol12,13-bis[(2',4'-difluorophenyl)acetate] was obtained. The structure wasconfirmed by NMR and mass spectral analysis.

EXAMPLE 86

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-(2'-hydroxyethylamino)phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(ii) 20-deoxy-20-[(2'S)-1'-hydroxyprop-2'-yl)amino]phorbol 12-myristate13-acetate;

(iii) 20-deoxy-20-(2'-hydroxymethylpiperidin-1-yl)phorbol12-(4'-biphenyl)acetate 13-acetate; and

(iv) 20-deoxy-20-[(2'-carboxyethyl)amino]phorbol12-[(2',2'-dimethyl)(2",4"-difluorophenyl)acetate 13-butyrate.

EXAMPLE 87 20-Deoxy-20-(imidazol-1'-yl)phorbol12,13-Bis[(2',4'-difluorophenyl)acetate] and20-Deoxy-20-(imidazol-2'-yl)phorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

Twelve mg of imidazole was added to 100 mg of 20-deoxy-20-bromophorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 1.5 mL of acetonitrilefollowed by 11 μL of pyridine 10 min later. After 2 h another 10 mg ofimidazole was added. Three hours later a drop of pyrollidinyl pyridinewas added. After 1.5 h the mixture was partitioned between ethyl acetateand phosphate buffer (pH 2). The organic layer was washed with phosphatebuffer (pH 8), dried over sodium sulfate and concentrated in vacuo. Theresidue was chromatographed [silica; methylene chloride/tetrahydrofuran(65:35)] to afford 33 mg of20-deoxy-20-(imidazol-1'-yl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] and 13 mg of20-deoxy-20-(imidazol-2'-yl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structures were confirmedby NMR and mass spectral analysis.

EXAMPLE 88 20-Deoxy-20-(4'-dimethylaminopyridinium-1'-yl)phorbol12-Myristate 13-Acetate, Bromide (Iodide) salt

A solution of 75 mg of 20-deoxy-20-bromophorbol 12-myristate 13-acetateand 1 mL of ethylene glycol in 1 mL of acetone was treated with 15 mg ofpotassium iodide and 30 mg of 4-dimethylaminopyridine at roomtemperature. After 30 min the mixture was partitioned between ethylacetate and phosphate buffer (pH 2). The organic layer was washed withphosphate buffer (pH 8), dried over sodium sulfate and concentrated invacuo. After purification by preparative liquid chromatography [silica;methylene chloride/methanol(50:50)]20-deoxy-20-(4'-dimethylaminopyridinium-1'-yl)phorbol12-myristate 13-acetate, bromide (iodide) salt, was obtained. It did notcrystallize.

EXAMPLE 89 20-Deoxy-20-[(3'-hydroxymethylphenyl)aminophorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

A total of 99 mg of 3-aminobenzyl alcohol was added over 1.5 h to 95 mgof 20-deoxy-20-oxophorbol 12,13-bis[(2',4'-difluorophenyl)acetate] in 3mL of acetonitrile/10% aq. acetic acid (9:1). To this mixture was thenadded 23 mg of sodium cyanoborohydride, and, after 25 min, phosphatebuffer (pH 2) was added. This solution was extracted with ethyl acetateand the organic layer washed with phosphate buffer (pH 8), filteredthrough N/N/S and concentrated in vacuo. Chromatographic purification[silica; hexane/ethyl acetate (55:45)] afforded 70 mg of20-deoxy-20-[(3'-hydroxymethylphenyl)amino]phorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and mass spectral analysis.

EXAMPLE 90

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-[(4'-(2'-hydroxyethyl)phenyl)amino]phorbol12,13-didecanoate;

(ii) 20-deoxy-20-[(3'-methoxycarbonylmethylphenyl)amino]phorbol12-myristate 13-acetate;

(iii) 20-deoxy-20-[(3'-cyanophenyl)amino]phorbol 12-(3'-phenoxy)benzoate13-butyrate;

(iv) 20-deoxy-20-[(4'-carboxamidophenyl)amino]phorbol12-(3',5'-difluorophenyl)acetate 13-(3',4'-difluorobenzoate);

(v) 20-deoxy-20-[(3'-acetylphenyl)amino]phorbol12,13-bis[(2',4'-dichlorophenyl)acetate];

(vi) 20-deoxy-20-[(3'-dihydroxyboranylphenyl)amino]phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(vii) 20-deoxy-20-[(4'-oxoarsinylphenyl)amino]phorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(viii) 20-deoxy-20-[(4'-dihydroxyphosphonylphenyl)amino]phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; and

(ix) 12,20-dideoxy-20-[(3'-hydroxymethylphenyl)amino]phorbol13-(2',4'-difluorophenyl)acetate.

EXAMPLE 91 20-O-(2'-Hydroxyethyl)phorbol 12-Myristate 13-Acetate

A solution of 75 mg of 20-deoxy-20-bromophorbol 12-myristate 13-acetateand 1 mL of ethylene glycol in 1 mL of acetone was treated with 15 mg ofpotassium iodide and 30 mg of 2,6-di-t-butyl-4-methylpyridine at roomtemperature in the dark and under a nitrogen atmosphere for 12 days. Itwas then partitioned between ethyl acetate and phosphate buffer (pH 2).The organic layer was washed with phosphate buffer (pH 8), filteredthrough N/N/S and concentrated in vacuo. After purification bypreparative liquid chromatography [silica; hexane/ethyl acetate (50:50)]28.5 mg of 20-O-(2'-hydroxyethyl)phorbol 12-myristate 13-acetate wasobtained. The structure was confirmed by NMR and high resolution massspectral analysis.

EXAMPLE 92

In a similar manner the following compounds are prepared:

(i) 20-O-(4'-hydroxy-2'-butynyl)phorbol 12,13bis[(2',4'-difluorophenyl)acetate];

(ii) 20-O-(4'-hydroxy-2'-butenyl)phorbol 12-(4'-biphenyl)acetate13-acetate;

(iii) 20-O-acetonylphorbol12-[(2',2'-dimethyl)(2",4"-difluorophenyl)acetate]13-butyrate;

(iv) 20-O-(2'-propyl)phorbol 12-myristate 13-acetate; and

(v) 20-O-(3'-hydroxyprop-1'-yl)phorbol 12-myristate 13-acetate.

EXAMPLE 93 20-O-(4'-Hydroxyphenoxy)carbonylphorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 8.26 g of hydroquinone and 6.3 g of imidazole in 80 mLof pyridine was added 20.6 g of t-butyldiphenylchlorosilane in 20 mL ofpyridine. After 1.5 h the mixture was partitioned between water andethyl acetate. The organic layer was washed twice with phosphate buffer(pH 2) and once each with phosphate buffer (pH 8) and brine before beingfiltered through N/N/S and concentrated in vacuo. Preparative liquidchromatography [silica; hexane/ethyl acetate (85:15)] afforded 14.4 g of4-(t-butyldiphenylsilyloxy)phenol. The structure was confirmed by itsNMR spectrum.

To a solution of 46 mg of 1,1'-carbonyldiimidazole in 0.5 mLnitromethane was added 65 μL of methyl triflate followed by 100 mg of4-(t-butyldiphenylsilyloxy)phenol in 1.4 mL of nitromethane. A total of1.9 mL of this solution was added to a mixture of 100 mg of phorbol12,13-bis[(2',4'-difluorophenyl)acetate] and approximately 10 mg of4-dimethylaminopyridine in 500 μL of anhydrous tetrahydrofuran. After4.5 h this mixture was partitioned between ethyl acetate and water. Theorganic layer was filtered through N/N/S, concentrated in vacuo andsubjected to preparative liquid chromatography [silica; hexane/ethylacetate (80:20)] to afford 142 mg of20-O-[4'-(t-butyldiphenylsilyloxy)phenoxy]carbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate]. A solution of this material in1 mL tetrahydrofuran was treated with 160 μL of 1.0M tetrabutylammoniumfluoride in tetrahydrofuran. After 14 min the mixture was partitionedbetween ethyl acetate and phosphate buffer (pH 8). The organic layer wasfiltered through N/N/S and concentrated in vacuo. After preparativeliquid chromatography [silica; hexane/ethyl acetate (65:35)] 108 mg of20-O-(4'-hydroxyphenoxy)carbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate] was obtained. The structure wasconfirmed by NMR and mass spectral analysis.

EXAMPLE 94

In a similar manner the following compounds are prepared:

(i) 20-O-(4'-hydroxyphenoxy)carbonylphorbol 12,13-didecanoate;

(ii) 20-O-(4'-hydroxyphenoxy)carbonyl-12-deoxyphorbol 13-decanoate;

(iii) 20-O-(4'-hydroxyphenoxy)carbonyl-12-deoxyphorbol13-(2',4'-difluorophenyl)acetate;

(iv) 20-O-(3'-hydroxyphenoxy)carbonylphorbol 12-myristate 13-acetate;

(v) 20-O-(3'-hydroxyphenoxy)carbonylphorbol 12-(3'-phenoxy)benzoate13-butyrate;

(vi) 20-O-(4'-hydroxyphenoxy)carbonylphorbol12-(3',5'-difluorophenyl)acetate 13-(3',4'-difluorobenzoate);

(vii) 20-O-(2'-hydroxyphenoxy)carbonylphorbol12-(3',5'-difluorocinnamate) 13-[3'-(2",4"-difluorophenyl)propionate];

(viii) 20-O-phenoxycarbonylphorbol12,13-bis[(2',4'-dichlorophenyl)acetate];

(ix) 20-O-(2'-hydroxyphenoxy)carbonylingenol 3-benzoate;

(x) 20-O-(3'-hydroxyphenoxy)carbonylresiniferonol9,13,14-orthophenylacetate;

(xi)3-[(4'-hydroxy)phenoxycarbonyloxymethyl]-1,6-dioxo-2,5-dioxacyclotetracosane;

(xii) 30-O-(4'-hydroxyphenoxy)carbonyldebromoaplysiatoxin 20-acetate;and

(xiii) 26-O-(4'-hydroxyphenoxy)carbonylbryostatin 1.

EXAMPLE 95 Phorbol 12-Myristate 13-Acetate20-(4'-Fluoro-3'-nitrophenyl)carbamate

To a solution of 100 mg of phorbol 12-myristate 13-acetate in 1 mL oftetrahydrofuran was added, 73 mg of4-fluoro-3-nitrophenyl isocyanate, 5mg of dibutyltin dilaurate and 25 mg of 4-dimethylaminopyridine. After 3h 100 μL of methanol was added to the reaction mixture. After removal ofthe solvents under a nitrogen stream, the mixture was subjected topreparative liquid chromatography [silica; hexane/iso-propyl alcohol(86:14)] to afford 71 mg of phorbol 12-myristate 13-acetate20-(4'-fluoro-3'-nitrophenyl)carbamate.

EXAMPLE 96

In a similar manner the following compounds are prepared:

(i) phorbol 12-myristate 13-acetate 20-ethylcarbamate;

(ii) phorbol 12-myristate 13-acetate 20-n-propylcarbamate;

(iii) phorbol 12-myristate 13-acetate 20-n-butylcarbamate;

(iv) phorbol 12,13-didecanoate 20-(3'-trifluoromethyl)phenylcarbamate;

(v) phorbol 12,13-bis[(2',4'-difluorophenyl)acetate]20-methylcarbamate;

(vi) 12-deoxyphorbol 13-decanoate 20-(3'-methoxy)phenylcarbamate;

(vii) 12-deoxyphorbol 13-(2',4'-difluorophenyl)acetate20-n-propylcarbamate;

(viii) phorbol 12-(3'-phenoxy)benzoate 13-butyrate20-(4'-ethoxycarbonyl)phenylcarbamate;

(ix) phorbol 12-(3',5'-difluorophenyl)acetate13-(3',4'-difluorobenzoate) 20-ethoxycarbonylmethylcarbamate;

(x) ingenol 3-benzoate 20-methylcarbamate;

(xi) resiniferonol 9,13,14-orthophenylacetate 20-ethylcarbamate;

(xii) mezerein 20-methylcarbamate;

(xiii)3-[(N-methylamino)carbonyloxymethyl]-1,6-dioxo-2,5-dioxacyclotetracosane;

(xiv) debromoaplysiatoxin 20-acetate 30-methylcarbamate; and

(xv) bryostatin 1 26-methylcarbamate.

EXAMPLE 97 Phorbol 12-Myristate 13-Acetate 20-Methylthiocarbamate

To a solution of 100 mg of phorbol 12-myristate 13-acetate in 1 mL oftetrahydrofuran was added, over a six hour period, a total of 180 mg ofmethyl isothiocyanate, 105 mg of dibutyltin dilaurate and 125 mg of4-dimethylaminopyridine. After 4 days the reaction mixture was subjectedto preparative liquid chromatography [silica; hexane/ethyl acetatemixtures] to afford 4 mg of phorbol 12-myristate 13-acetate20-methylthiocarbamate.

EXAMPLE 98

In a similar manner the following compounds are prepared:

(i) phorbol 12,13-didecanoate 20-allylthiocarbamate;

(ii) phorbol12,13-bis[(2',4'-difluorophenyl)acetate]20-(tetrahydrofuran-2'-yl)methylthiocarbamate;

(iii) 12-deoxyphorbol 13-(2',4'-difluorophenyl)acetate20-(3'-methoxy)propylthiocarbamate; and

(iv) phorbol 12-(3'-phenoxy)benzoate 13-butyrate20-phenethylthiocarbamate.

EXAMPLE 99 Phorbol 12-Myristate 13-Acetate 20-Carbamate

To a solution of 100 mg of phorbol 12-myristate 13-acetate in 2 mL ofmethylene chloride was added 25 mg of chlorosulfonyl isocyanate in 200μL of methylene chloride. After 3 h a few hundred μL of phosphate buffer(pH 2) was added. After 20 min pH 8 buffer was added and the mixtureextracted with ethyl acetate. The organic layer was filtered throughN/N/S and concentrated in vacuo. After preparative liquid chromatography[silica; hexane/tetrahydrofuran (65/35)], 57 mg of phorbol 12-myristate13-acetate 20-carbamate was obtained.

EXAMPLE 100

In a similar manner the following compounds are prepared:

(i) phorbol 12,13-bis[(2',4'-difluorophenyl)acetate]20-carbamate;

(ii) phorbol 12,13-didecanoate 20-carbamate;

(iii) phorbol 12-(3'-phenoxy)benzoate 13-butyrate 20-carbamate; and

(iv) 12-deoxyphorbol 13-(2',4'-difluorophenyl)acetate 20-carbamate.

EXAMPLE 10120-Deoxy-20-(2'-hydroxyethylsulfonyl)-3-deoxo-3-hydroximinophorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a solution of 100 mg of 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 1 mL of pyridine was added214 mg of hydroxylamine hydrochloride. After heating at 53° C. for 8 hthe mixture was cooled and partitioned between phosphate buffer (pH 2)and ethyl acetate. The organic layer was washed with phosphate buffer(pH 8), dried over sodium sulfate and concentrated in vacuo. Preparativeliquid chromatography of the residue [silica; methylenechloride/tetrahydrofuran (92.5:7.5)] afforded 62 mg of20-deoxy-20-(2'-hydroxyethylsulfonyl)-3-deoxo-3-hydroximinophorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and mass spectral analysis.

EXAMPLE 102

In a similar manner the following compounds are prepared:

(i) 6-deshydroxymethyl-6-carboxy-3-deoxo-3-hydroximinophorbol12,13-bis[(2',4'-difluorophenyl)acetate]; and

(ii) 20-deoxy-20-(2'-hydroxyethylthio)-3-deoxo-3-hydroximinophorbol12,13-bis[3'-(pentafluorophenyl)propionate].

EXAMPLE 10320-Deoxy-20-(2'-hydroxyethylthio)-3-deoxo-3-beta-hydroxyphorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a mixture of 25 mg of 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] and 15 mg of cerium (III)chloride in 0.5 mL of methanol was added approximately 3 mg of sodiumborohydride. After 8 min the mixture was partitioned between phosphatebuffer (pH 2) and ethyl acetate. The organic layer was washed withphosphate buffer (pH 8), dried over sodium sulfate and concentrated by astream of nitrogen. Purification by preparative liquid chromatography[silica; hexane/ethyl acetate (50:50)] afforded 13 mgof20-deoxy-20-(2'-hydroxyethylthio)-3-deoxo-3-beta-hydroxyphorbol12,13-bis[(2',4'-difluorophenyl)acetate]. The structure was confirmed byNMR and high resolution mass spectral analysis.

EXAMPLE 104

In a similar manner the following compounds are prepared:

(i) 3-deoxo-3-beta-hydroxyphorbol12-[4'-(9",10"-dihydrophenanthren-2")butyrate]; and

(ii) 6-deshydroxymethyl-6-carboxy-3-deoxo-3-beta-hydroxyphorbol12,13-bis[(2',4'-difluorophenyl)acetate].

EXAMPLE 105 20-Deoxyphorbol 12-Myristate 13-Acetate and12-beta-Myristoyloxy-13-acetoxy-4,9-dihydroxy-1,6(20)-tigladien-3-oneand 20-Deoxy-20-[20'-deoxyphorbol-20'-yl (12'-myristate13'-acetate)]phorbol 12-Myristate 13-Acetate

Extensive further purification of the crude residue from Example 15 bypreparative liquid chromatography afforded 20-deoxyphorbol 12-myristate13-acetate,12-beta-myristoyloxy-13-acetoxy4,9-dihydroxy-1,6(20)-tigladien-3-one and20-deoxy-20-[20'-deoxyphorbol-20'-yl (12'-myristate 13'-acetate)]phorbol12-myristate 13-acetate. These structures were confirmed by NMR and highresolution mass spectral analysis.

EXAMPLE 106 6-Deshydroxymethyl-6-[2'-(methoxycarbonyl)-(E)-vinyl]phorbol12-Myristate 13-Acetate

A solution of 100 mg of20-deoxy-20-oxophorbol 12-myristate 13-acetateand 90 mg of methyl triphenylphosphoranylideneacetate in 4 mL of toluenewas allowed to stir for 18 hours at room temperature. Afterconcentration in vacuo, the residue was purified by preparative liquidchromatography [silica; methylene chloride/tetrahydrofuran (96:4)] toyield 116 mg of6-deshydroxymethyl-6-[2'-(methoxycarbonyl)-(E)-vinyl]phorbol12-myristate 13-acetate as an oil.

EXAMPLE 107 20-Deoxyphorbol 12-Myristate 13-Acetate 20-Sulfonic Acid,Triethylamine Salt

A mixture containing 100 mg of20-deoxy-20-chlorophorbol 12-myristate13-acetate, 22 mg of sodium sulfite, 20 mg of sodium iodide and 0.6mmoles of acetic acid in about 11 mL of ethanol/water (approximately1:1) was stirred for a total of 4 hours alternating between ambient and60° C. After that time the mixture was concentrated in vacuo andpartitioned between ethyl acetate and brine to afford a residue whichwas purified by liquid chromatography [silica; methylenechloride/methanol (90:10)] to yield 82 mg of 20-deoxyphorbol-20-sulfonicacid 12-myristate 13-acetate. This acid was converted to itstriethylamine salt by the addition of 24 μL of triethylamine in t-butylmethyl ether followed by concentration in vacuo.

EXAMPLE 108 20-Deoxy-20-trimethylphosphoniumphorbol12,13-Bis[(2',4'-difluorophenyl)acetate]Bromide

To solution of approximately 50 mg of 20-deoxy-20-bromophorbol12,13-bis[(2',4'-difluorophenyl)acetate] in 1 mL of dry acetonitrile isadded 100 μL of 1M trimethylphosphine in toluene. After a period of timethis mixture is concentrated under a nitrogen stream and the residue issubjected to chromatographic purification to afford20-deoxy-20-trimethylphosphoniumphorbol12,13-bis[(2',4'-difluorophenyl)acetate]bromide.

EXAMPLE 109 20-Deoxy-20-trimethylsilylphorbol12,13-Bis[(2',4'-difluorophenyl)acetate]

To a suspension of 50 mg of 20-deoxy-20-bromophorbol12,13-bis[(2',4'-difluorophenyl)acetate] and 40 mg of powdered zinc(less than 325 mesh) in 500 μL of anhydrous tetrahydrofuran was added 15μL trimethylchlorosilane. After stirring overnight at room temperaturethe reaction mixture was treated with another 50 μL of silane andsonicated for 5 h. The mixture is then partitioned between ethyl acetateand phosphate buffer (pH 8) and chromatographed in the usual way toobtain 20-deoxy-20-trimethylsilylphorbol12,13-bis[(2',4'-difluorophenyl)acetate].

EXAMPLE 110

In a similar manner the following compounds are prepared:

(i) 20-deoxy-20-diphenyloxophosphinylphorbol12,13-bis[(2',4'-difluorophenyl)acetate];

(ii) 20-deoxy-20-trimethylsilylphorbol 12-myristate 13-acetate; and

(iii) 20-deoxy-20-diphenyloxophosphinylphorbol 12-myristate 13-acetate.

EXAMPLE 111 Phorbol 12-[4'-(9",10"-Dihydrophenanthrene-2")butyrate]

A solution of 2.95 g of 4-(9',10'-dihydrophenanthrene-2')butyric acidand 1.13 g of dicyclohexylcarbodiimide in 20 mL of tetrahydrofuran waswarmed briefly with hot air and then stirred for 5 min at roomtemperature. The mixture was then filtered and concentrated in vacuo toobtain 2.79 g of the 4-(9',10'-dihydrophenanthrene-2')butyric anhydride.This anhydride was mixed with 1.6 g of20-O-[diphenyl(4'-methoxyphenyl)methyl]phorbol13-O-isopropyldimethylsilyl then 15 mL of pyridine was added. After 18 hof stirring under nitrogen 100 mg of 4-dimethylaminopyridine was addedfollowed 7 h later with 400 mg of dicyclohexylcarbodiimide. Afteranother 22 h at room temperature approximately 10 mL of methanol wasadded followed by the concentration of the mixture in vacuo. The residuewas partitioned between ethyl acetate and phosphate buffer (pH 2) andthe organic layer washed with phosphate buffer (pH 8), filtered throughN/N/S and again concentrated in vacuo. Purification by preparativeliquid chromatography [silica; hexane/ethyl acetate (75:25)] affordedslightly impure 20-O-[diphenyl(4'-methoxyphenyl)methyl]phorbol12-[4'-(9",10"-dihydrophenanthrene-2")butyrate]13-O-isopropyldimethylsilyl.

The entire sample of the above compound was dissolved in 35 mL oft-butylmethyl ether and treated with 150 mL of 90% acetic acid in water.After 70 min the mixture was concentrated in vacuo and partitionedbetween ethyl acetate and phosphate buffer (pH 8) followed byconcentration of the organic layer in vacuo. Preparative liquidchromatography [silica; methylene chloride/methanol (97:3)] afforded 890mg of phorbol 12-[4'-(9",10"-dihydrophenanthrene-2")butyrate].

EXAMPLE 112 20-O-[Diphenyl(4'-methoxyphenyl)methyl]phorbol12-[3',5'-Bis(trifluoromethyl)phenylcarbamate]13-Methoxyacetate

To a solution of 396 mg of20-O-[diphenyl(4'-methoxyphenyl)methyl]phorbol 13-methoxyacetate, 70 mgof 4-dimethylaminopyridine and 72 mg of dibutyltin dilaurate in 5 mL oftetrahydrofuran was added 220 mg of 3,5-bis(trifluoromethyl)phenylisocyanate in 1 mL tetrahydrofuran. After 7 h added another 100 mg ofthe isocyanate. After stirring for another 12 h at room temperature thereaction mixture was partitioned between ethyl acetate/hexane and water.The organic layer was filtered through N/N/S and concentrated in vacuoto afford 872 mg of crude 20-O-[diphenyl(4'-methoxyphenyl)methyl]phorbol12-[3',5'-bis(trifluoromethyl)phenylcarbamate]13-methoxyacetate.

EXAMPLE 113

In a similar manner the following compounds are prepared:

(i) 20-O-[diphenyl(4'-methoxyphenyl)methyl]phorbol12-n-octadecylcarbamate 13-methoxyacetate; and

(ii) 20-O-[diphenyl(4'-methoxyphenyl)methyl]phorbol12-(pentafluorophenyl)acetate 13-methoxyacetate (the acylating agent waspentafiuorophenylacetyl fluoride).

EXAMPLE 114 Phorbol 12-[3',5'-Bis(trifluoromethyl)phenylcarbamate]

To a solution of 174 mg of20-O-[diphenyl(4'-methoxyphenyl)methyl]phorbol12-[3',5'-bis(trifluoromethyl)phenylcarbamate]13-methoxyacetate in 5 mLof methanol was added approximately 20 mg of sodium carbonate. After 4.5h the mixture was partitioned between water and ethyl acetate. Theorganic layer was filtered through N/N/S and concentrated in vacuo. Theresulting residue was dissolved in 2.7 mL dry methylene chloride and 0.7mL of absolute ethanol and treated with 50 μL of trifluoroacetic acid.After 1.5 h the mixture was partitioned between ethyl acetate andphosphate buffer (pH 8). The organic layer was filtered through N/N/Sand concentrated in vacuo. Preparative liquid chromatography [silica;hexane/ethyl acetate (50:50)] afforded 27 mg of phorbol12-[3',5'-bis(trifluoromethyl)phenylcarbamate]. The structure wasconfirmed by NMR and high resolution mass spectral analysis.

EXAMPLE 115

In a similar manner the following compounds are prepared:

(i) phorbol 12-n-octadecylcarbamate; and

(ii) phorbol 12-(pentafluorophenyl)acetate.

EXAMPLE 116 Demonstration of Anti-HIV Activity of20-Deoxy-20-chlorophorbol 12,13-bis[(2',4'-difluorophenyl)acetate]

Human peripheral blood lymphocytes were isolated from the buffy coatfractions of blood donations. The lymphocytes were then stimulated with5 micrograms/ml of phytohemagglutinin for 48 hours. Prior to infectionwith HIV, the lymphocytes were washed and resuspended in mitogen-freemedium. On day 0 the cells were infected with HIV and were cultured forfour days in the presence or absence of graded concentrations of20-deoxy-20-chlorophorbol 12,13-bis[(2',4'-difluorophenyl)acetate]. Ondays 3 and 4 the supernatant levels of total viral RNA and viral coreprotein p24 were determined at each drug concentration and dose-responsecurves were used to determine the concentration of drug giving 50%inhibition of production of viral RNA and of p24 core protein. At fourdays these ED₅₀ values were less than 100 pmolar for both RNA and p24.

EXAMPLE 117 Demonstration of Anti-HIV Activity of Diterpenoid Phorboids

In a manner similar to Example 116, the anti-HIV ED₅₀ values for RNA[drug concentration is in brackets] for the following diterpenoidcompounds were determined from dose-response curves or by estimationfrom one or more experimental drug concentrations:

(i) phorbol 12-[4'-(9",10"-dihydrophenanthrene-2")butyrate] [450nmolar];

(ii) 20-deoxy-20-bromophorbol 12,13-bis[(2',4'-difluorophenyl)acetate][less than 1 nmolar];

(iii) 20-deoxy-20-chlorophorbol 12,13-bis[(pentafluorophenyl)acetate][less than 1 nmolar];

(iv) 20-deoxy-20-cyanophorbol 12-myristate 13-acetate [1.5 μmolar];

(v) phorbol 12-myristate 13-acetate20-(4'-fluoro-3'-nitrophenyl)carbamate [4 μmolar];

(vi) 20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] [1.0 μmolar];

(vii) 20-deoxy-20-(2',3'-dihydroxypropylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate][3μmolar];

(viii) 20-deoxy-20-hydroximinophorbol12,13-bis[(2',4'-difluorophenyl)acetate] [5.5 μmolar];

(ix) 20-deoxy-3-deoxo-3,20-bis(hydroximino)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] [45 nmolar];

(x) 6-deshydroxymethyl-6-carboxyphorbol12,13-bis[(2',4'-difluorophenyl)acetate] [0.1-1 μmolar];

(xi) 6-deshydroxymethyl-6-methoxycarbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate] [1.6 μmolar];

(xii) 20-deoxy-20-(2'-carboxyethylthio)phorbol12,13-bis(2',4'-difluorophenylacetate) [4.5 μmolar];

(xiii) 20-deoxy-20-(2'-hydroxyethylsulfinyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] [1.6 μmolar];

(xiv) 20-deoxy-20-(2'-hydroxyethylsulfonyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] [5.0 μmolar];

(xv) 20-deoxy-20-(2'-hydroxyethylsulfinyl)phorbol 12-myristate13-acetate [1.1 μmolar];

(xvi) 20-deoxy-20-(2'-hydroxyethylsulfonyl)phorbol 12-myristate13-acetate [500 nmolar];

(xvii) 20odeoxy-20-[(3'-hydroxymethylphenyl)amino]phorbol12,13-bis[(2',4'-difluorophenyl)acetate] [180 nmolar];

(xviii) 20-deoxy-20-azidophorbol12,13-bis[(2',4'-difluorophenyl)acetate] [65 nmolar];

(xix) 20-deoxy-20-aminophorbol 12,13-bis[(2',4'-difluorophenyl)acetate][100 nmolar];

(xx) 20-O-(4'-hydroxyphenoxy)carbonylphorbol12,13-bis[(2',4'-difluorophenyl)acetate] [less than 10 nmolar];

(xxi) 20-deoxy-20-propylthiophorbol12,13-bis[(2',4'-difluorophenyl)acetate] [10 nmolar];

(xxii) 20-deoxy-20-propylsulfinylphorbol12,13-bis[(2',4'-difluorophenyl)acetate] [500 nmolar];

(xxiii) 20-deoxy-20-(2'-mercaptoethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate] [50 nmolar]; and

(xxiv) 6-deshydroxymethyl-6-cyanophorbol12,13-bis[(2',4'-difluorophenyl)acetate] [less than 100 nmolar].

(xxv) 20-deoxy-20-fluorophorbol 12,13-bis[(pentafluorophenyl)acetate] [1nmolar];

EXAMPLE 118 Demonstration of Anti-HIV Activity of Indolactam-TypePhorboids

In a manner similar to Example 116, the anti-HIV ED₅₀ values for RNA[drug concentration is in brackets] for the following indolactam-typecompounds were determined from dose-response curves or by estimationfrom one or more experimental drug concentrations; data giving percentinhibition of viral RNA production at selected concentrations appear inparentheses:

(i) rac-14-O-(N-methyl)carbamoylindolactam V (52% inhibition at 10 μM);

(ii)rac-14-O-(N-methyl)carbamoyl-1-N-(2'-triphenylphosphonium)ethylindolactamV, methanesulfonate salt (82% inhibition at 10 μM);

(iii) 14-O-(N-methyl)carbamoyl-7-octyl-(9S,12S)-indolactam V (87%inhibition at 10 μM);

(iv) 9-deshydroxymethyl-9-[N-(2'-glucosyl)]carboxamidoindolactam V (35%inhibition at 10 μM);

(v) 9-deshydroxymethyl-9-[N-(2',3'dihydroxypropyl)]carboxamidoindolactamV (22% inhibition at 10 μM);

(vi) 9-deshydroxymethyl-9-ethoxycarbonylindolactam V (47% inhibition at10 μM);

(vii) 1-N-hydroxymethyl-9-deshydroxymethyl-9-ethoxycarbonylindolactam V(31% inhibition at 10 μM);

(viii) 9-deshydroxymethyl-9-(2',3'-dihydroxy)propyloxycarbonylindolactamV (20% inhibition at 10 μM);

(ix) 14-deoxy-14-(2'-hydroxyethylthio)indolactam V [1μM]; and

(x) 14-deoxy-14-(2'-hydroxyethylthio)-7-octyl-(9S,12S)-indolactam V [100nM].

EXAMPLE 119 Demonstration of Anti-H IV Activity of Diacylglycerol-TypePhorboids

In a manner similar to Example 116, the anti-HIV ED₅₀ values for RNA[drug concentration is in brackets] for the followingdiacylglycerol-type compounds were determined from dose-response curvesor by estimation from one or more experimental drug concentrations; datagiving percent inhibition of viral RNA production at selectedconcentrations appear in parentheses:

(i) 3-[(2'-hydroxyethylthio)methyl]-1,6-dioxo-2,5-dioxacyclotetracosane[10 nmolar].

EXAMPLE 120 Demonstration of Anti-melanoma Activity

Human RPMI-7272 melanoma cells were grown in the standard culture mediumunder normal incubation conditions. On day 1 the cells were cultured inthe absence (control) or presence of graded concentrations (3-100 μg/ml)of 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-myristate 13-acetate inseparate tubes. On day 4 after 72 h of exposure the number of cells ineach tube was measured and the number of cell doublings determined. Thedrug treated tubes were compared to the control tube. The ID₅₀ (theconcentration of drug required to inhibit cell doublings by 50%) was 42μM.

In a like manner the anti-melanoma activity of phorbol12-[4'-(9",10"-dihydrophenanthrene-2")butyrate] was determined, ID₅₀ 12μM.

EXAMPLE 121 Demonstration of Anti-leukemic Activity

HL-60 promyelocytic leukemia cells were cultured in RPMI 1640 mediumsupplemented with 10% fetal bovine serum. Cells (7,500) were seeded into96-well microtiter plates and incubated overnight. Serial dilutions of20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-myristate 13-acetate(dissolved in DMSO and then diluted with culture medium) were added tothe wells on day 1. The plates were incubated for 8 days to allow thecontrol cultures to undergo at least 3 cell divisions. The cell growthwas monitored by using the calorimetric MTT (tetrazolium) assay[Mosmann, T., J. Immunol. Meth., 65 55-63 (1983)]. After the incubationperiod, the cells were washed with phosphate-buffered saline in themicrotiter plate. DMSO was then added to each well and the dish was puton a shaker for 20 min. The optical density was measured at 540 nm andcompared using the formula: (OD Test-OD Start)/(OD Control-ODStart)×100. The IC₅₀ (the drug concentration which leads to 50% of cellsper well compared to control cultures (100%) at the end of theincubation period) is 5.4 μM. In a like manner, the anti-leukemicactivities of the following other compounds were demonstrated:

(i) phorbol 12-myristate 13-acetate 20-methylcarbamate (IC₅₀ =2.6 μM);

(ii) 20-deoxy-20-cyanophorbol 12-myristate 13-acetate (IC₅₀ =5 μM);

(iii)12-beta-myristoyloxy-13-acetoxy-4,9-dihydroxy-7-hydroxymethyl-1,6(20)-tigladien-3-one(IC₅₀ =5.2 μM);

(iv) phorbol 12-(pentafluorophenyl)acetate (IC₅₀ =50.8 μM); and

(v) phorbol 12-[4'-(9", 10"-dihydrophenanthrene-2")butyrate] (IC₅₀ =1.8μM).

EXAMPLE 122 Demonstration of Anti-cancer Activity

T-24 human bladder carcinoma cells were cultured in Eagle's minimalessential medium supplemented with 5% fetal bovine serum. Cells (1,000)were seeded into 96-well microtiter plates and incubated overnight.Serial dilutions of 20-deoxy-20-(2'-hydroxyethylthio)phorbol12-myristate 13-acetate (dissolved in DMSO and then diluted with culturemedium) were added to the wells on day 1. The plates were incubated for5-6 days to allow the control cultures to undergo at least 3 celldivisions. After the incubation period, the cells were fixed withglutaraldehyde, washed with water and stained with 0.05% methylene blue.After washing the dye was eluted with 3% HCl. The optical density perwell was measured at 665 nm and compared using the formula: (OD Test-ODStart)/(OD Control-OD Start)×100. The IC₅₀ (the drug concentration whichleads to 50% of cells per well compared to control cultures (100%) atthe end of the incubation period) is 13 μM.

In a like manner, the anti-cancer activities of the following othercompounds were demonstrated:

(i) phorbol 12-myristate 13-acetate 20-methylcarbamate (IC₅₀ =6.3 μM);

(ii) 20-deoxy-20-cyanophorbol 12-myristate 13-acetate (IC₅₀ =2 μM);

(iii)12-beta-myristoyloxy-13-acetoxy4,9-dihydroxy-7-hydroxymethyl-1,6(20)-tigiadien-3-one(IC₅₀ =5.5 μM); and

(iv) phorbol 12-[4'-(9",10"-dihydrophenanthrene-2")butyrate] (IC₅₀ =4.0μM).

EXAMPLE 123 Gelatin Capsules

Gelatin capsules containing the following ingredients are prepared:

    ______________________________________                                        rac-14-O-(N-methyl)carbamoyl-1-N-                                                                    125 mg                                                 (2'-triphenylphosphonium)ethyl-                                               indolactam V, methanesulfonate salt                                           lactose                300 mg                                                 talc                    15 mg                                                 ______________________________________                                    

The finely powdered ingredients are blended together. The mixture isused to fill hard shell two-piece gelatin capsules of a suitable size ata net fill weight of 440 mg.

EXAMPLE 124 Injectable Solution

A suspension (1.0 ml) suitable for intramuscular injection is preparedfrom the following ingredients:

    ______________________________________                                        14-deoxy-14-(3'-hydroxypropylthio)indolactam V                                                            20    mg                                          polyethylene glycol 3350    29    mg                                          polysorbate 80              2     mg                                          monobasic sodium phosphate  6.8   mg                                          dibasic sodium phosphate    1.4   mg                                          benzyl alcohol              9     mg                                          water for injection to make 1.0   ml                                          ______________________________________                                    

The materials are mixed, homogenized and filled into 1 ml ampuls whichare sealed.

EXAMPLE 125 Topical Gel

An illustrative composition for a topical gel is the following:

    ______________________________________                                        20-deoxy-20-cyanophorbol 12-myristate                                                                20 mg                                                  13-acetate                                                                    hydroxypropylcellulose 60 mg                                                  ethyl alcohol          920 mg                                                 ______________________________________                                    

The materials are mixed, homogenized and filled into containers eachholding 1 gram of gel.

EXAMPLE 126 Topical Solution

An illustrative composition for a topical solution is the following:

    ______________________________________                                        14-deoxy-14-[(2',3'-hydroxy)propylthio]-                                                              20        mg                                          teleocidin B-1                                                                propylene glycol        300       mg                                          citric acid             20        mg                                          SD alcohol 40-2         to make 1.0                                                                             ml                                          ______________________________________                                    

The materials are mixed, homogenized and filled into squeezable plasticcontainers each holding 1 milliliter of solution.

Equivalents

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, many equivalents to the specificembodiments of the invention described herein. Such equivalents areintended to be encompassed by the following claims.

We claim:
 1. A compound, in the form of an individual isomer, an isomermixture, a racemate or optical antipode, or a pharmaceuticallyacceptable salt thereof, of the formula:

    P--G

wherein P represents a radical, formally derived from a parent compoundof the diterpene-type phorboid class, which compound:a. binds reversiblyor irreversibly to a diacylglycerol-type receptor; and/or b. activatesany form of the enzyme protein kinase C; and c. contains anhydroxymethyl or 1-hydroxyethyl group bonded to a carbon atom; andwherein G is any group of 55 or fewer atoms selected from carbon,hydrogen, oxygen, nitrogen, halogen, sulfur, phosphorus, silicon,arsenic, boron and selenium either i) singly or doubly bonded to thecarbon atom of the parent compound in place of the hydroxymethyl or1-hydroxyethyl group; or ii) singly or doubly bonded to a carbon atomimmediately adjacent to the carbon atom to which the hydroxymethyl or1-hydroxyethyl group is bound in the parent compound; and wherein thehydroxymethyl or 1-hydroxyethyl group of the parent compound is absentor has been replaced by G; provided that: for all P, when G is bonded tothe equivalent of phorbol's carbon 6, G may not be --CH₃, --CH═O, --CH₂O-R_(es) wherein R_(es) is an acyl group, or --CH₂ O--R_(et) whereinR_(et) is a (i) C₁ -C₆ hydrocarbon radical, (ii) a substituted orunsubstituted triphenylmethyl group, (iii) a (C₁ -C₆ linear or branchedalkyl)_(n) (phenyl)_(3-n) silyl group, wherein n is 0-3, or is (iv)--C(CH₃)₂ O--, --C(═O)O-- or --B(C₆ H₅)O-- linked via the oxygen atom tocarbon 5 of P, and G may not be --CH═NNHR_(h) wherein R_(h) is a phenylring with or without substituents; P--G may not include12-beta-13-alpha-diacetoxy derivatives of compounds having the exact20-carbon tigliane or the exact 19-carbon 20-nor-tigliane skeleton; P--Gmay not comprise 20-deoxy-20-chlorocrotophorbolone nor20-deoxy-20-chlorophorbol 12,13-diesters wherein the ester groups areboth selected from saturated or unsaturated alkanoyl or are bothbenzoyl; if G is ═CH₂ bonded to carbon 6 of P,P may not carry a ringformed by --OC(CH₃)₂ O--bonded in the beta configuration to carbons 3and 4 or a trimethylsilyloxy at carbon 4; if P has an ingenol skeletonwith a rearranged A-ring, G may not consist or --CH₂ --OC(═O)NHC₆ H₅ ;and, P--G may not comprise 6-deshydroxymethyl-6-carboxyphorbol12,13-didecanoate, 6-deshydroxymethyl-6-carboxyphorbol 12,13-diacetate,20-chloro-12,13,20-trideoxy-3-deoxo-3α-trimethylsilyloxy-4-O-trimethylsilyl-1,2-dihydro-2(19)-didehydrophorbol,20-iodo-9,12,13,20-tetradeoxy-3-deoxo-3α-benzoyloxy-4-O-trimethylsilyl-1,2,6,7-tetrahydro-2(19)-didehydro-6α,9α-oxidophorbol,20-O-benzyl-9,12,13-trideoxy-3-deoxo-3α-benzoyloxy-4-O-trimethylsilyl-1,2,6,7-tetrahydro-2(19)-didehydro-6α,9α-oxidophorbolor1,2-didehydro-6-deshydroxymethyl-2-desmethyl-4,9,12,13-tetradeoxy-4,7-dimethylphorbol.2. A compound, in the form of an individual isomer, an isomer mixture, aracemate or optical antipode, or a pharmaceutically acceptable saltthereof, of the formula:

    P.sub.o --S.sub.o --E.sub.o

wherein P_(o) represents a radical, formally derived from a parentcompound of the diterpene-type phorboid class, which compound:a. bindsreversibly or irreversibly to a diacylglycerol-type receptor; and/or b.activates any form of the enzyme protein kinase C; and c. contains anhydroxymethyl or 1-hydroxyethyl group bonded to a carbon atom; andwherein S_(o) --E_(o) represents a substituent which is either i) singlyor doubly bonded to the carbon atom of the parent compound in place ofthe hydroxymethyl or 1-hydroxyethyl group; or ii) singly or doublybonded to a carbon atom immediately adjacent to the carbon atom to whichthe hydroxymethyl or 1-hydroxyethyl group is bound in the parentcompound; and wherein S_(o) is an organic substituted or unsubstituted,saturated, unsaturated and/or aromatic, straight or branched, acyclic,ring-containing and/or ring-carrying chain of atoms which separatesP_(o) and E_(o) by a linear count of at least two but not more than 12atoms and contains and/or carries not more than 9 heteroatoms selectedfrom oxygen, nitrogen, silicon, sulfur, phosphorus, arsenic, boron andselenium, and not more than 16 halogen atoms; provided that the totalnumber of atoms does not exceed about 35; and wherein E_(o) compriseshydrogen, halogen or a saturated or singly or multiply unsaturated groupcontaining up to 15 carbon atoms and optionally containing 1 to 12halogen atoms and/or 1 to 6 heteroatoms selected from oxygen, nitrogen,silicon, sulfur, phosphorus, arsenic, boron and selenium; or whereinS_(o) E_(o) taken together may be a hydrogen, halogen, thionic sulfuratom or ketonic oxygen atom or a hydroxy, amine, or thiol group singlyor doubly bonded to the carbon atom of the parent compound P_(o) inplace of the hydroxymethyl or 1-hydroxyethyl group; provided that: forall P_(o), S_(o) E_(o) taken together, when bonded to carbon 6 may notbe --CH₂ O--R_(et) wherein R_(et) is (i) an acyl group, (ii) a C₁ -C₆hydrocarbon radical, (iii) a substituted or unsubstitutedtriphenylmethyl group, (iv) a (C₁ -C₆ linear or branched alkyl)_(n)(phenyl)_(3-n) silyl, wherein n is 0-3, or is (v) --C(CH₃)₂ O--,--C(═O)O-- or --B(C₆ H₅)O-- linked via the oxygen atom to the βconfiguration of carbon 5 of Po, and S_(o) E_(o) may not be--CH═NNHR_(h) wherein R_(h) is a phenyl ring with or withoutsubstituents; P_(o) --S_(o) --E_(o) may not include 12-β-13-α-diacetoxyderivatives of compounds having the exact 20-carbon tigliane or theexact 19-carbon 20-nor-tigliane skeleton; P_(o) --S_(o) --E_(o) may notcomprise 20-deoxy-20-chlorocrotophorbolone nor 20-deoxy-20-chlorophorbol12,13-diesters wherein the ester groups are both selected from saturatedor unsaturated alkanoyl or are both benzoyl; if P_(o) has an ingenolskeleton with a rearranged A-ring, S_(o) E_(o) may not consist of --CH₂--OC(═O)NHC₆ H₅ ; and, P_(o) --S_(o) --E_(o) may not comprise6-deshydroxymethyl-6-carboxyphorbol 12,13-didecanoate,6-deshydroxymethyl-6-carboxyphorbol 12,13-diacetate,20-O-benzyl-9,12,13-trideoxy-3-deoxo-3α-benzyloxy-4-O-trimethylsilyl-1,2,6,7-tetrahydro-2(19)-didehydro-6α,9α-oxidophorbolor 1,2-didehydro-6-deshydroxymethyl-2-desmethyl-4,9,12,13-tetradeoxy-4,7-dimethylphorbol.
 3. A compound of the formula:

    P.sub.1 --S.sub.1 --E.sub.1

wherein P₁ is a radical of the formula: ##STR40## in the form of anindividual isomer, an isomer mixture, a racemate or optical antipode, ora pharmaceutically acceptable salt thereof, wherein A¹ and A² may beindependently selected from hydrogen, halogen and a straight chain orbranched chain, cyclic or acyclic, saturated, unsaturated and/oraromatic carbon- and/or heteroatom-containing substituent having notmore than 34 carbon atoms, not more than 24 halogen atoms and not morethan 9 heteroatoms selected from oxygen, nitrogen, silicon, phosphorusand sulfur, or A¹ and A² taken together may complete a 5- or 6-membered,saturated or unsaturated, carbocyclic or heterocyclic ring, optionallysubstituted by halogen(s) and/or by 1-8 straight chain or branchedchain, cyclic or acyclic, saturated, unsaturated, and/or aromaticcarbon- and/or heteroatom-containing groups, which halogens and groupstaken together contain a total of not more than 30 carbon atoms, notmore than 24 halogen atoms and not more than 9 heteroatoms selected fromoxygen, nitrogen, silicon, phosphorus and sulfur; A³ is a three atomchain which carries S₁ E₁ and completes a 7-membered saturated orunsaturated carbocyclic ring optionally substituted by 1-6 substituentsindependently selected from halogen(s) and straight chain or branchedchain, cyclic or acyclic, saturated, unsaturated, and/or aromaticcarbon- and/or heteroatom-containing groups, which halogens and groupstaken together, excluding S₁ E₁, contain not more than 12 carbon atoms,not more than 8 halogen atoms, and not more than 5 heteroatoms selectedfrom oxygen, nitrogen, silicon, phosphorus and sulfur; provided that,including S₁ E₁, the middle carbon atom of A³ is not substituted byhydroxymethyl or hydroxyethyl; A⁴ completes a 6- or 7-memberedcarbocyclic or heterocyclic ring (connected in the beta configuration toeither carbon atom 9 or 10), optionally substituted by halogen(s) and/orby 1-10 straight chain or branched chain, cyclic or acyclic, saturated,unsaturated and/or aromatic carbon- and/or heteroatom-containing groups,the group or groups optionally completing 1-3 additional rings throughbonds among themselves and/or 1-5 additional rings when taken togetherwith A¹, A², a ring formed by A¹ and A² together, and/or a bond tocarbon atom 9, which halogens and groups taken together, include notmore than 50 carbon atoms, not more than 24 halogen atoms, and not morethan 15 heteroatoms selected from oxygen, nitrogen, silicon, phosphorusand sulfur; carbon atom 9 may be optionally bound to a substituent by asingle or double bond; J¹ is selected from hydrogen, fluoro, chloro,hydroxy, amino, mono- or di-loweralkylamino, methyl, ethyl, vinyl,ethynyl, propargyl, cyano, methoxy, ethoxy, trifluoromethyl,2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, acetoxy, propanoyloxy,acetyl, propanoyl, hydroxyacetyl, 2-hydroxypropanoyloxy,3-hydroxypropanoyl, acetamido, propanamido, hydroxyacetamido,2-hydroxypropanamido, or 3-hydroxypropanamido (each of which must besituated in the beta configuration), or J¹ taken together with A¹, A²,A³ or a ring formed by A¹ together with A² completes a 3- to 7-memberedsubstituted or unsubstituted carbocyclic or heterocyclic ring, thesubstituents of which contain not more than 15 carbon atoms, not morethan 10 halogens, and not more than 8 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur; J² is selected from hydrogen,methyl, ethyl, hydroxymethyl, hydroxyethyl, vinyl, ethynyl, allyl,propargyl, n-propyl and isopropyl; wherein S₁ is a chain of atoms of theformula: ##STR41## wherein a, b, d, e, and g may independently be from 0to 3; c and f may independently be 0 or 1; the sum of (a+b+c+d+e+f+g) isat least 1 but not more than 12; and if c and f are both 1, then the sumof (d+e) must be at least 1; R₁ ¹ through R₁ ¹² may be the same ordifferent and each may be hydrogen, halogen or a saturated or singly ormultiply unsaturated, straight or branched, acyclic substituentcontaining not more than 20 carbon atoms, not more than 16 halogenatoms, and not more than 6 heteroatoms selected from oxygen, nitrogen,and sulfur, silicon and phosphorus; R₁ ¹ or R₁ ² may optionally be anadditional bond completing an unsaturated linkage to P₁ ; one or two ofthe substituents R₁ ¹ -R₁ ¹² may optionally comprise the same ordifferent values of G¹, as defined below; R₁ ¹¹ or R₁ ¹² may optionallybe an additional bond to E₁, thereby completing an unsaturated linkage;one of the substituents R₁ ¹ -R₁ ¹² may be linked to either the atom inP₁ that carries the S₁ chain or to an atom in P₁ adjacent thereto, toform a saturated, unsaturated or aromatic, carbocyclic or heterocyclic4-8 membered ring optionally containing 1-4 identical or different ringhetero members selected from O, S, SO, SO₂, CO, ═N--, and NH, the ringbeing optionally substituted on any carbon and/or NH members, by 1-8identical or different substituents selected from halogen, hydroxy,methoxy, ethoxy, methyl, ethyl, cyano, azide, nitro, hydroxymethyl,1-hydroxyethyl, 2-hydroxy-2-propyl, CF₃, OCF₃, SH, SCH₃, SOCH₃, SCF₃,COOH, COOCH₃, COCH₃, CH═O, acetoxy, amino, mono- or dialkylaminototalling 1-4 carbon atoms inclusive, acetamido, N-methylacetamido,carboxamido, N-alkylated carboxamido containing 1-4 carbon atomsinclusive, hydroxyacetyl and hydroxyacetoxy; X and X' are as definedbelow; provided that, for all substituents R₁ ¹ through R₁ ¹² and allconstituents X and X', taken together, but excluding any atoms of P₁ :the total of carbon atoms is 25 or less; the total of halogen atoms is16 or less; the total of oxygen atoms is 6 or less; the total ofnitrogen atoms is 4 or less; the total of sulfur atoms is 3 or less; thetotal of oxygen, nitrogen, silicon, phosphorus and sulfur atoms togetheris 8 or less; the total of --OH groups is 3 or less; the total of --NH₂groups is 2 or less; the total of --SH groups is 2 or less; the total of--OH, --SH, and --NH₂ groups together is 4 or less; X and X' may be thesame or different and are selected from: ##STR42## wherein R_(X) ¹,R_(X) ², R_(X) ¹¹ and R_(X) ¹² may independently be hydrogen; R_(X) ¹through R_(X) ¹² may be the same or different and each may be an acyclicsubstituent containing 1-20 carbon atoms, not more than 16 halogenatoms, and not more than 6 heteroatoms selected from oxygen, nitrogen,and sulfur, such that for any substituent the oxygen atoms total 4 orless, the nitrogen atoms total 4 or less, and the sulfur atoms total 2or less; R_(X) ⁴, R_(X) ⁵, R_(X) ¹¹ and R_(X) ¹² may independently behydroxy; Q and Q' are as defined below; R_(X) ¹ may optionally representan additional bond to P₁, thus completing an unsaturated linkage; one ortwo of the substituents R_(X) ¹ -R_(X) ¹² may optionally comprise thesame or different values of G¹, as defined below; and one of thesubstituents R_(X) ¹ -R_(X) ¹² may be linked to either the atom in P₁that carries the chain containing X, X', and/or X" or to an atom in P₁adjacent thereto, to form a saturated, unsaturated or aromatic,carbocyclic or heterocyclic 4-8 membered ring optionally containing 1-4other identical or different hetero ring members selected from O, S, SO,SO₂, CO, ═N--, and NH, the ring being optionally substituted on itscarbon and/or NH members by 1-8 identical or different substituentsselected from halogen, hydroxy, methoxy, ethoxy, methyl, ethyl, cyano,azide, nitro, hydroxymethyl, 1-hydroxyethyl, 2-hydroxy-2-propyl, CF₃,OCF₃, SH, SCH₃, SOCH₃, SCF₃, COOH, COOCH₃, COCH₃, CH═O, acetoxy, amino,mono- or dialkylamino totalling 1-4 carbon atoms inclusive, acetamido,N-methylacetamido, carboxamido, N-alkylated carboxamido containing 1-4carbon atoms inclusive, hydroxyacetyl and hydroxyacetoxy; Q and Q'independently may be:

    ═O═S═N--R.sub.Q.sup.1 or ═N--O--R.sub.Q.sup.2

wherein R_(Q) ¹ and R_(Q) ² may be the same or different and each may behydrogen or a saturated or singly or multiply unsaturated, straight orbranched, acyclic substituent containing 1-20 carbon atoms, not morethan 16 halogen atoms, and not more than 6 heteroatoms selected fromoxygen, nitrogen, and sulfur, in which the oxygen atoms total 4 or less,the nitrogen atoms total 4 or less, and the sulfur atoms total 2 orless, the heteroatoms being preferably situated in functional groupsselected from hydroxy, amino, thiol, nitro, azide, ether, thioether,aldehyde, keto, carboxy, ester, amide, cyano, nitroguanidine, andcyanoguanidine; R_(Q) ¹ and/or R_(Q) ² may optionally comprise the sameor different values for G¹, as defined below; R_(Q) ¹ may be linked toeither the atom in P₁ that carries the chain containing Q and/or Q' orto an atom in P₁ adjacent thereto, to form a 4-8 membered ring definedas for R₁ ¹ -R₁ ¹² -containing ring above; G¹ comprises a groupcontaining 1-3 fused or separate, saturated, unsaturated, or aromatic,carbocyclic or heterocyclic 4-8 membered rings, each ring optionallycontaining 1-4 identical or different hetero ring members selected fromO, S, SO, SO2, CO, ═N--, and NH, each ring being optionally substitutedon its carbon and/or NH members by 1-8 identical or differentsubstituents selected from halogen, hydroxy, methoxy, ethoxy, methyl,ethyl, cyano, azide, nitro, hydroxymethyl, 1-hydroxyethyl,2-hydroxyethyl, 2-hydroxy-2-propyl, CF₃, OCF₃, SH, SCH₃, SOCH₃, SCF₃,COOH, COOCH₃, COCH₃, CH═O, acetoxy, amino, mono- or dialkylaminototalling 1-4 carbon atoms inclusive, acetamido, N-methylacetamido,carboxamido, N-alkylated carboxamido containing 1-4 carbon atomsinclusive, hydroxyacetyl and hydroxyacetoxy; and wherein for G¹ theoptional second and third rings may be fused to the first ring and/or toone another or may be separate rings connected to one another and/or tothe atom bearing G¹ by a single or double bond or by an interveningsubstituted or unsubstituted, linear or branched, saturated orunsaturated chain containing not more than 8 carbon atoms, not more than8 halogens, and not more than 4 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur; T³ may also be hydrogen orhalogen; T⁵ -T^(5") are independently selected from hydrogen andhydroxy; T⁵ may also be halogen; R_(E) ¹ is selected from hydrogen,halogen, hydroxy, nitro, nitroso, cyano, azide, --NH₂, --NH--OH, --SH,--O--NH₂, --NH--NH₂, --T¹ --C(═T²)--T³, --C(═T²)--T³, --SiT⁵ T^(5')T^(5"), --T¹ --S(═O)(═T²)--T⁴, --S(═O)(═T²)--T⁴, --T¹ --P(--T⁴)--T^(4'),--P(--T⁴)--T^(4'), --T¹ --P(═T^(2'))(--T³)--T⁴, and--P(═T^(2'))(--T³)--T⁴ ; R_(E) ² and R_(E) ³ are individually selectedfrom hydrogen, --C(═T²)--T³, cyano, nitro, azide, halogen and a C₁ -C₁₅straight or branched chain, saturated, unsaturated and/or aromaticsubstituent optionally containing not more than 10 halogen atoms and notmore than 4 heteroatoms selected from oxygen, nitrogen and sulfur; ifR_(E) ¹ is cyano or C(═T²)--T³, then R_(E) ² or R_(E) ³ may optionallybe selected from --SiT⁵ T^(5') T^(5"), --T¹ --P(═T^(2'))(--T³)--T⁴, and--P(═T^(2'))(--T³)--T⁴ ; and R_(E) ⁴ and R_(E) ⁵ are individuallyselected from hydrogen, halogen, cyano, nitro, C(═T²)--T³, --T¹--C(═T²)--T³, --CR_(E) ¹ R_(E) ² R_(E) ³, --SiT⁵ T^(5') T^(5"),--S(═O)(═T²)--T⁴, and --P(═T^(2'))(--T³)--T⁴ ; provided that: for allP₁, S₁ E₁ taken together, when bonded to carbon 6, may not be --CH₂O--R_(et) wherein R_(et) is a C₁ -C₆ hydrocarbon radical, a substitutedor unsubstituted triphenylmethyl group, a C₁ -C₆ linear or branchedalkyl)_(n) (phenyl)_(3-n) silyl, wherein n is 0-3, or is --C(CH₃)₂ O--,--C(═O)O-- or --B(C₆ H₅)O-- linked via the oxygen atom to carbon 5 of P₁; P₁ --S₁ --E₁ may not include 12--β13-α-diacetoxy derivatives ofcompounds having the exact 20-carbon tigliane or the exact 19-carbon20-nor-tigliane skeleton; P₁ --S₁ --E₁ may not comprise20-O-benzyl-9,12,13-trideoxy-3-deoxo-3α-benzoyloxy-4-O-trimethylsilyl-1,2,6,7-tetrahydro-2(19)-didehydro-6α,9α-oxidophorbol;and, the total wherein E₁ is selected from ═O, ═S, ═NH, ═NOR_(E) ⁸ whereR_(E) ⁸ is hydrogen or a C₁ -C₈ normal or branched alkyl radical,═N--NH₂, hydrogen, halogen, --OH, --SH, --NH₂, --NH--NH₂, --N₃, --CN,--NO, --NO₂, --NHOH, --ONH₂, or is selected from: ##STR43## wherein T¹is selected from --O--, --S--, and --NH--; T² is selected from ═O , ═S,and ═N--R_(E) ⁶, in which R_(E) ⁶ may be hydrogen, hydroxy, cyano, ornitro; T^(2') is selected from ═O and ═S; T³, T⁴ and T^(4') areindependently selected from --OH, --NH₂, --SH, --N₃, --NH--NH₂, and--NH--OR_(E) ⁷, in which R_(E) ⁷ may be hydrogen, C₁₋₃ alkyl, or C₁₋₃acyl; P₁ ring skeleton may not comprise any of the following six minimumframeworks ##STR44## with or without additional ring(s) appendedthereto.
 4. A compound of claim 3 wherein S₁ and E₁ taken togethercomprise:(i) --P(═O)(R_(a) ^(a))OH, --P(═S)(R_(a) ^(a))OH, --CH═NOCH₃,--CH═N(→O)CH₃, --C(CH₃)═NOH, in which R_(a) ^(a) is C₁₋₁₂ linear orbranched, saturated, unsaturated and/or aromatic hydrocarbon optionallysubstituted by not more than 16 halogens; or (ii) --CH₂ -- or--CH(CH₃)--, to either of which is bonded --SR_(a) ^(a), --S(═O)R_(a)^(a), --SCH₂ CH₂ OH, --S(═O)--CH₂ CH₂ OH, --S(CH₂)₃ OH, --S(CH₂)₄ OH,--SCH₂ CH₂ SH[, --M--C(═T²)--M'--R_(a) ^(a), --OCH₂ C(═O)CH₃, --OCH₂C(═NOH)CH₃, the o-, m- or p-isomer of --M--C(═T²)--M'--C₆ H₄ --T³, theo-, m- or p-isomer of --M--C(═T²)--M'--C₆ H₄ CH₂ --T³, or-imidazol-2-yl].
 5. A compound of claim 3 wherein P₁ is P_(1R),##STR45## wherein carbons (1 and 2) or (2 and 3) may be joined by adouble bond; carbons (5 and 6) or (6 and 7) may be joined by a doublebond;S₁ E₁ may be bonded to carbon 5, 6 or 7; R_(A1) represents not morethan 8 identical or different substituents bonded independently viasingle and/or double bonds to carbons 1, 2 and/or 3, which substituentsmay independently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing groups, which halogens and groups taken togethercontain a total of not more than 30 carbon atoms, not more than 24halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur; R_(A3) represents not morethan 6 identical or different substituents bonded independently viasingle and/or double bonds to carbons 5, 6 and/or 7, which substituentsmay independently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated, and/or aromatic carbon-and/or heteroatom-containing groups, which halogens and groups takentogether contain not more than 12 carbon atoms, not more than 8 halogenatoms, and not more than 5 heteroatoms selected from oxygen, nitrogen,silicon, phosphorus and sulfur; and R_(A4) represents not more than 10identical or different substituents bonded independently via single ordouble bonds to carbons 9, 11, 12, 13 and/or 14, which substituents mayindependently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing groups, the group or groups optionally completing1-3 additional rings through bonds among themselves and/or 1-5additional rings when taken together with the 5-membered ring and itssubstituent(s) R_(A1), which halogens and groups taken together, includenot more than 50 carbon atoms, not more than 24 halogen atoms, and notmore than 15 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur.
 6. A compound of claim 5 wherein carbons 13 and14 of P_(1R) carry a substituted or unsubstituted cyclopropyl ring,forming P_(1P) ##STR46## wherein the R_(A5) and R_(A6) radicals mayindependently be hydrogen, halogen and/or a straight chain or branchedchain, cyclic or acyclic, saturated, unsaturated and/or aromatic carbon-and/or heteroatom-containing group, which halogens and groups takentogether contain a total of not more than 30 carbon atoms, not more than24 halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur.
 7. A compound of claim 6wherein S₁ and E₁ taken together comprise:(i) --P(═O)(R_(a) ^(a))OH,--P(═S)(R_(a) ^(a))OH, --CH═NOCH₃, --CH═N(→O)CH₃, --C(CH₃)═NOH, in whichR_(a) ^(a) is C₁₋₁₂ linear or branched, saturated, unsaturated and/oraromatic hydrocarbon optionally substituted by not more than 16halogens; or (ii) --CH₂ -- or --CH(CH₃)--, to either of which is bonded--SR_(a) ^(a), --S(═O)R_(a) ^(a), --SCH₂ CH₂ OH, --S(═O)--CH₂ CH₂ OH,--S(CH₂)₃ OH, --S(CH₂)₄ OH, --SCH₂ CH₂ SH.
 8. A compound of claim 6wherein the P_(1P) carbon 2 carries a methyl group, J¹ is hydroxy,carbon 9 carries a hydroxy group in the alpha configuration, carbons 10and 14 carry alpha hydrogens, carbon 11 carries a methyl group in thealpha configuration and R_(A5) and R_(A6) are both methyl, formingP_(1PP) ##STR47##
 9. A compound of claim 8 selected from the groupconsisting of:(i) 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-butyrate;(ii) 20-deoxy-20-(2'-hydroxyethylthio)phorbol 12,13-dibutyrate; (iii)20-deoxy-20-(2'-hydroxyethylthio)phorbol 12-myristate 13-acetate; (iv)20-deoxy-20-[(2'-hydroxyethyl)methylamino]phorbol 12-myristate13-acetate; (v) 20-deoxy-20-(3'-hydroxypropylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; (vi)20-deoxy-20-N-methylaminophorbol 12-myristate 13-acetate; (vii) phorbol12-myristate 13-acetate 20-cholinephosphate; (viii)20-deoxy-20-(2',3'-dihydroxypropylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; (ix)20-deoxy-20-[2'-(formylamino)ethylthio]phorbol 12-myristate 13-acetate;(x) 20-deoxy-20-[2'-(methylsulfonylamino)ethylthio]phorbol 12-myristate13-acetate; (xi) 20-deoxy-20-propylthiophorbol12,13-bis[(2',4'-difluorophenyl)acetate]; (xii)20-deoxy-20-(2'-mercaptoethylthio)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; (xiii)20-deoxy-20-(2'-hydroxyethylthio)phorbol12,13-[O,O'-bis(isopropyldimethylsilyl)]; (xiv)20-deoxy-20-(2'-carboxyethylthio)phorbol 12-(4'-biphenyl)acetate13-[3'-(3",5"-difluorophenyl)propionate]; (xv)20-deoxy-20-(2'-hydroxyethylsulfinyl)phorbol12,13-bis[(pentafluorophenyl)acetate]; (xvi)20-deoxy-20-(propylsulfinyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; (xvii)20-deoxy-20-(propylsulfonyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; and (xviii)20-O-(2'-hydroxyethyl)phorbol 12-myristate 13-acetate.
 10. A compound ofclaim 8 wherein S₁ E₁ taken together comprise a saturated normal orbranched C₂ -C₁₀ alkyl group containing one hydroxy group separated fromP₁ by not more than four carbon atoms.
 11. A compound of claim 10selected from the group consisting of:(i)20-deoxy-20-(1'-hydroxyethyl)phorbol 12-myristate 13-acetate; (ii)20-deoxy-20-(2'-hydroxyethyl)phorbol 12-myristate 13-acetate; (iii)20-deoxy-20-(2'-hydroxyethyl)phorbol12,13-bis[(2',4'-difluorophenyl)acetate]; and (iv)12,20-dideoxy-20-(2'-hydroxyethyl)phorbol 13-angelate.
 12. A compound ofclaim 5 wherein an orthoester structure is bonded to P_(1R) via oneorthoester oxygen atom in the alpha configuration to carbon 9 and viatwo orthoester oxygen atoms in the alpha configuration to any two ofcarbons 12-14, forming P_(1RR) ##STR48## wherein R_(A7) representshydrogen, halogen or a straight chain or branched chain, cyclic oracyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing group, the group optionally completing 1additional ring to carbon 1, which group includes not more than 30carbon atoms, not more than 16 halogen atoms, and not more than 9heteroatoms selected from oxygen, nitrogen, silicon, phosphorus andsulfur.
 13. A compound of claim 12 wherein S₁ and E₁ taken togethercomprise:(i) --P(═O)(R_(a) ^(a))OH, --P(═S)(R_(a) ^(a))OH, --CH═NOCH₃,--CH═N(→O)CH₃, --C(CH₃)═NOH, in which R_(a) ^(a) is C₁₋₁₂ linear orbranched, saturated, unsaturated and/or aromatic hydrocarbon optionallysubstituted by not more than 16 halogens; or (ii) --CH₂ -- or--CH(CH₃)--, to either of which is bonded --SR_(a) ^(a), --S(═O)R_(a)^(a), --SCH₂ CH₂ OH, --S(═O)--CH₂ CH₂ OH, --S(CH₂)₃ OH, --S(CH₂)₄ OH,--SCH₂ CH₂ SH.
 14. A compound of claim 12 wherein the P_(1RR) carbon 2carries a methyl group, J¹ is hydroxy, carbon 10 carries a hydrogen atomin the alpha configuration, carbon 11 carries a methyl group in thealpha configuration and carbon 13 carries an isopropenyl group, formingP_(1RRR) ##STR49##
 15. A compound of claim 14 selected from the groupconsisting of:(i) 20-deoxy-20-(2'-hydroxyethylthio)resiniferonol9,13,14-orthophenylacetate; (ii)20-deoxy-20-(2'-carboxyethylthio)resiniferonol9,13,14-orthophenylacetate; and (iii)20-deoxy-20-(3'-hydroxypropylthio)mezerein.
 16. A compound of claim 3wherein P₁ is P_(1I) ##STR50## wherein R_(A2I) represents not more than8 identical or different substituents bonded independently via singleand/or double bonds to carbons 1, 2 and/or 3, which substituents mayindependently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing groups, which halogens and groups taken togethercontain a total of not more than 30 carbon atoms, not more than 24halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur;R_(A3I) represents not morethan 6 identical or different substituents bonded independently viasingle and/or double bonds to carbons 5, 6 and/or 7, which substituentsmay independently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated, and/or aromatic carbon-and/or heteroatom-containing groups, which halogens and groups takentogether contain not more than 12 carbon atoms, not more than 8 halogenatoms, and not more than 5 heteroatoms selected from oxygen, nitrogen,silicon, phosphorus and sulfur; and R_(A4I) represents not more than 10identical or different substituents bonded independently via single ordouble bonds to carbons 9, 11, 12,13 and/or 14, which substituents mayindependently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing groups, the group or groups optionally completing1-3 additional rings through bonds among themselves and/or 1-5additional rings when taken together with the 5-membered ring and itssubstituent(s) R_(A2I), which halogen and groups taken together, includenot more than 50 carbon atoms, not more than 24 halogen atoms, and notmore than 15 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur.
 17. A compound of claim 16 wherein carbons 13 and14 of P_(1I) carry a substituted or unsubstituted cyclopropyl ring,forming P_(1IN) ##STR51## wherein the R_(A5I) and R_(A6I) radicals mayindependently be hydrogen, halogen and/or a straight chain or branchedchain, cyclic or acyclic, saturated, unsaturated and/or aromatic carbon-and/or heteroatom-containing group, which halogens and groups takentogether contain a total of not more than 30 carbon atoms, not more than24 halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur.
 18. A compound of claim 17wherein S₁ and E₁ taken together comprise:(i) --P(═O)(R_(a) ^(a))OH,--P(═S)(R_(a) ^(a))OH, [--P(═O)(OR_(a) ^(a))OH, --P(═S)(OR_(a) ^(a))OH,--CH═NOH,]--CH═NOCH₃, --CH═N(→O)CH₃, --C(CH₃)═NOH, in which R_(a) ^(a)is C₁₋₁₂ linear or branched, saturated, unsaturated and/or aromatichydrocarbon optionally substituted by not more than 16 halogens; or (ii)--CH₂ -- or --CH(CH₃)--, to either of which is bonded --SR_(a) ^(a),--S(═O)R_(a) ^(a), --SCH₂ CH₂ OH, --S(═O)--CH₂ CH₂ OH, --S(CH₂)₃ OH,--S(CH₂)₄ OH, --SCH₂ CH₂ SH.
 19. A compound of claim 17 wherein theP_(1IN) carbon 2 carries a methyl group, J¹ is hydroxy, carbon 11carries a methyl group in the alpha configuration, carbon 14 carries analpha hydrogen and R_(A5I) and R_(A6I) are both methyl, forming P_(1INL)##STR52##
 20. A compound of claim 19 selected from the group consistingof:(i) 20-deoxy-20-(n-propylthio)ingenol 3-benzoate; and (ii)20-deoxy-20-(n-propylthio)ingenol 3-(2',4'-difluorobenzyl)carbamate. 21.A composition, comprising:a physiologically acceptable pharmaceuticalcarrier; and a compound, in a quantity of between about 0.0001-1000 mgper unit dosage, in the form of an individual isomer, an isomer mixture,a racemate or optical antipode, or a pharmaceutically acceptable saltthereof, of the formula:

    P--G

wherein P represents a radical, formally derived from a parent compoundof the diterpene-type phorboid class, which compound:a. binds reversiblyor irreversibly to a diacylglycerol-type receptor; and/or b. activatesany form of the enzyme protein kinase C; and c. contains anhydroxymethyl or 1-hydroxyethyl group bonded to a carbon atom;andwherein G is any group of 55 or fewer atoms selected from carbon,hydrogen, oxygen, nitrogen, halogen, sulfur, phosphorus, silicon,arsenic, boron and selenium either i) singly or doubly bonded to thecarbon atom of the parent compound in place of the hydroxymethyl or1-hydroxyethyl group; or ii) singly or doubly bonded to a carbon atomimmediately adjacent to the carbon atom to which the hydroxymethyl or1-hydroxyethyl group is bound in the parent compound; and wherein thehydroxymethyl or 1-hydroxyethyl group of the parent compound is absentor has been replaced by G.
 22. A composition, comprising:aphysiologically acceptable pharmaceutical carrier; and a compound, in aquantity of between about 0.0001-1000 mg per unit dosage, in the form ofan individual isomer, an isomer mixture, a racemate or optical antipode,or a pharmaceutically acceptable salt thereof, of the formula:

    P.sub.o --S.sub.o --E.sub.o

wherein P_(o) represents a radical, formally derived from a parentcompound of the diterpene-type phorboid class, which compound:a. bindsreversibly or irreversibly to a diacylglycerol-type receptor; and/or b.activates any form of the enzyme protein kinase C; and c. contains anhydroxymethyl or 1-hydroxyethyl group bonded to a carbon atom; andwherein S_(o) --E_(o) represents a substituent which is either i) singlyor doubly bonded to the carbon atom of the parent compound in place ofthe hydroxymethyl or 1-hydroxyethyl group; or ii) singly or doublybonded to a carbon atom immediately adjacent to the carbon atom to whichthe hydroxymethyl or 1-hydroxyethyl group is bound in the parentcompound; and wherein S_(o) is a substituted or unsubstituted,saturated, unsaturated and/or aromatic, straight or branched, acyclic,ring-containing and/or ring-carrying chain of atoms which separatesP_(o) and E_(o) by a linear count of at least two but not more than 12atoms and contains and/or carries not more than 9 heteroatoms selectedfrom oxygen, nitrogen, silicon, sulfur, phosphorus, arsenic, boron andselenium, and not more than 16 halogen atoms; provided that the totalnumber of atoms does not exceed 35; and wherein E_(o) compriseshydrogen, halogen or a saturated or singly or multiply unsaturated groupcontaining up to 15 carbon atoms and optionally containing 1 to 12halogen atoms and/or 1 to 6 heteroatoms selected from oxygen, nitrogen,silicon, sulfur, phosphorus, arsenic, boron and selenium; or whereinS_(o) E_(o) taken together may be a hydrogen, halogen, thionic sulfuratom or ketonic oxygen atom or a hydroxy, amine, or thiol group singlyor doubly bonded to the carbon atom of the parent compound P_(o) inplace of the hydroxymethyl or 1-hydroxyethyl group.
 23. A composition,comprising:a physiologically acceptable pharmaceutical carrier; and acompound, in a quantity of between about 0.0001-1000 mg per unit dosage,of the formula:

    P.sub.1 --S.sub.1 --E.sub.1

wherein P₁ is a radical of the formula: ##STR53## in the form of anindividual isomer, an isomer mixture, a racemate or optical antipode, ora pharmaceutically acceptable salt thereof, wherein A¹ and A² may beindependently selected from hydrogen, halogen and a straight chain orbranched chain, cyclic or acyclic, saturated, unsaturated and/oraromatic carbon- and/or heteroatom-containing substituent having notmore than 34 carbon atoms, not more than 24 halogen atoms and not morethan 9 heteroatoms selected from oxygen, nitrogen, silicon, phosphorusand sulfur, or A¹ and A² taken together may complete a 5- or 6-membered,saturated or unsaturated, carbocyclic or heterocyclic ring, optionallysubstituted by halogen(s) and/or by 1-8 straight chain or branchedchain, cyclic or acyclic, saturated, unsaturated, and/or aromaticcarbon- and/or heteroatom-containing groups, which halogens and groupstaken together contain a total of not more than 30 carbon atoms, notmore than 24 halogen atoms and not more than 9 heteroatoms selected fromoxygen, nitrogen, silicon, phosphorus and sulfur; A³ is a three atomchain which carries S₁ E₁ and completes a 7-membered saturated orunsaturated carbocyclic ring optionally substituted by 1-6 substituentsindependently selected from halogen(s) and straight chain or branchedchain, cyclic or acyclic, saturated, unsaturated, and/or aromaticcarbon- and/or heteroatom-containing groups, which halogens and groupstaken together, excluding S₁ E₁, contain not more than 12 carbon atoms,not more than 8 halogen atoms, and not more than 5 heteroatoms selectedfrom oxygen, nitrogen, silicon, phosphorus and sulfur; provided that,including S₁ E₁, the middle carbon atom of A³ is not substituted byhydroxymethyl or hydroxyethyl; A⁴ completes a 6- or 7-memberedcarbocyclic or heterocyclic ring (connected in the beta configuration toeither carbon atom 9 or 10), optionally substituted by halogen(s) and/orby 1-10 straight chain or branched chain, cyclic or acyclic, saturated,unsaturated and/or aromatic carbon- and/or heteroatom-containing groups,the group or groups optionally completing 1-3 additional rings throughbonds among themselves and/or 1-5 additional rings when taken togetherwith A¹, A², a ring formed by A¹ and A² together, and/or a bond tocarbon atom 9, which halogens and groups taken together, include notmore than 50 carbon atoms, not more than 24 halogen atoms, and not morethan 15 heteroatoms selected from oxygen, nitrogen, silicon, phosphorusand sulfur carbon atom 9 may be optionally bound to a substituent by asingle or double bond; J¹ is selected from hydrogen, fluoro, chloro,hydroxy, amino, mono- or di- loweralkylamino, methyl, ethyl, vinyl,ethynyl, propargyl, cyano, methoxy, ethoxy, trifluoromethyl,2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, acetoxy, propanoyloxy,acetyl, propanoyl, hydroxyacetyl, 2-hydroxypropanoyloxy,3-hydroxypropanoyl, acetamido, propanamido, hydroxyacetamido,2-hydroxypropanamido, or 3-hydroxypropanamido (each of which must besituated in the beta configuration), or J¹ taken together with A¹, A²,A³ or a ring formed by A¹ together with A² completes a 3- to 7-memberedsubstituted or unsubstituted carbocyclic or heterocyclic ring, thesubstituents of which contain not more than 15 carbon atoms, not morethan 10 halogens, and not more than 8 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur; J² is selected from hydrogen,methyl, ethyl, hydroxymethyl, hydroxyethyl, vinyl, ethynyl, allyl,propargyl, n-propyl and isopropyl; wherein S₁ is a chain of atoms of theformula: ##STR54## wherein a, b, d, e, and g may independently be from 0to 3; c and f may independently be 0 or 1; the sum of(a+b+c+d+e+f+g) isat least 1 but not more than 12; and if c and f are both 1, then the sumof(d+e) must be at least 1; R₁ ¹ through R₁ ¹² may be the same ordifferent and each may be hydrogen, halogen or a saturated or singly ormultiply unsaturated, straight or branched, acyclic substituentcontaining not more than 20 carbon atoms, not more than 16 halogenatoms, and not more than 6 heteroatoms selected from oxygen, nitrogen,and sulfur, silicon and phosphorus; R₁ ¹ or R₁ ² may optionally be anadditional bond completing an unsaturated linkage to R₁ ; one or two ofthe substituents R₁ ¹ -R₁ ¹² may optionally comprise the same ordifferent values of G¹, as defined below; R₁ ¹¹ or R₁ ¹² may optionallybe an additional bond to E₁, thereby completing an unsaturated linkage;one of the substituents R₁ -R₁ ¹² may be linked to either the atom in P₁that carries the S₁ chain or to an atom in P₁ adjacent thereto, to forma saturated, unsaturated or aromatic, carbocyclic or heterocyclic 4-8membered ring optionally containing 1-4 identical or different ringhetero members selected from O, S, SO, SO₂, CO, ═N--, and NH, the ringbeing optionally substituted on any carbon and/or NH members, by 1-8identical or different substituents selected from halogen, hydroxy,methoxy, ethoxy, methyl, ethyl, cyano, azide, nitro, hydroxymethyl,1-hydroxyethyl, 2-hydroxy-2-propyl, CF₃, OCF₃, SH, SCH₃, SOCH₃, SCF3,COOH, COOCH₃, COCH₃, CH=0, acetoxy, amino, mono- or dialkylaminototalling 1-4 carbon atoms inclusive, acetamido, N-methylacetamido,carboxamido, N-alkylated carboxamido containing 1-4 carbon atomsinclusive, hydroxyacetyl and hydroxyacetoxy; X and X' are as definedbelow; provided that, for all substituents R₁ ¹ through R₁ ¹² and allconstituents X and X', taken together, but excluding any atoms of P₁ :the total of carbon atoms is 25 or less; the total of halogen atoms is16 or less; the total of oxygen atoms is 6 or less; the total ofnitrogen atoms is 4 or less; the total of sulfur atoms is 3 or less; thetotal of oxygen, nitrogen, silicon, phosphorus and sulfur atoms togetheris 8 or less; the total of --OH groups is 3 or less; the total of --NH₂groups is 2 or less; the total of --SH groups is 2 or less; the total of--OH, --SH, and --NH₂ groups together is 4 or less; X and X' may be thesame or different and are selected from: ##STR55## wherein R_(X) ¹,R_(X) ², R_(X) ¹¹ and R_(X) ¹² may independently be hydrogen; R_(X) ¹through R_(X) ¹² may be the same or different and each may be an acyclicsubstituent containing 1-20 carbon atoms, not more than 16 halogenatoms, and not more than 6 heteroatoms selected from oxygen, nitrogen,and sulfur, such that for any substituent the oxygen atoms total 4 orless, the nitrogen atoms total 4 or less, and the sulfur atoms total 2or less; R_(X) ⁴, R_(X) ⁵, R_(X) ¹¹ and R_(X) ¹² may independently behydroxy; Q and Q' are as defined below; R_(X) ¹ may optionally representan additional bond to P₁, thus completing an unsaturated linkage; one ortwo of the substituents R_(X) ¹ -R_(X) ¹² may optionally comprise thesame or different values of G¹, as defined below; and one of thesubstituents R_(X) ¹ -R_(X) ¹² may be linked to either the atom in P₁that carries the chain containing X, X', and/or X" or to an atom in P₁adjacent thereto, to form a saturated, unsaturated or aromatic,carbocyclic or heterocyclic 4-8 membered ring optionally containing 1-4other identical or different hetero ring members selected from O, S, SO,SO₂, CO, ═N--, and NH, the ring being optionally substituted on itscarbon and/or NH members by 1-8 identical or different substituentsselected from halogen, hydroxy, methoxy, ethoxy, methyl, ethyl, cyano,azide, nitro, hydroxymethyl, 1-hydroxyethyl, 2-hydroxy-2-propyl, CF₃,OCF₃, SH, SCH₃, SOCH₃, SCF₃, COOH, COOCH₃, COCH₃, CH═O, acetoxy, amino,mono- or dialkylamino totalling 1-4 carbon atoms inclusive, acetamido,N-methylacetamido, carboxamido, N-alkylated carboxamido containing 1-4carbon atoms inclusive, hydroxyacetyl and hydroxyacetoxy; Q and Q'independently may be:

    ═O═S═N--R.sub.Q.sup.1 or ═N--O--R.sub.Q.sup.2

wherein R_(Q) ¹ and R_(Q) ² may be the same or different and each may behydrogen or a saturated or singly or multiply unsaturated, straight orbranched, acyclic substituent containing 1-20 carbon atoms, not morethan 16 halogen atoms, and not more than 6 heteroatoms selected fromoxygen, nitrogen, and sulfur, in which the oxygen atoms total 4 or less,the nitrogen atoms total 4 or less, and the sulfur atoms total 2 orless, the heteroatoms being preferably situated in functional groupsselected from hydroxy, amino, thiol, nitro, azide, ether, thioether,aldehyde, keto, carboxy, ester, amide, cyano, nitroguanidine, andcyanoguanidine; R_(Q) ¹ and/or R_(Q) ² may optionally comprise the sameor different values for G¹, as defined below; R_(Q) ¹ may be linked toeither the atom in P₁ that carries the chain containing Q and/or Q' orto an atom in P₁ adjacent thereto, to form a 4-8 membered ring definedas for R₁ ¹ -R₁ ¹² -containing ring above; G¹ comprises a groupcontaining 1-3 fused or separate, saturated, unsaturated, or aromatic,carbocyclic or heterocyclic 4-8 membered rings, each ring optionallycontaining 1-4 identical or different hetero ring members selected fromO, S, SO, SO₂, CO, ═N--, and NH, each ring being optionally substitutedon its carbon and/or NH members by 1-8 identical or differentsubstituents selected from halogen, hydroxy, methoxy, ethoxy, methyl,ethyl, cyano, azide, nitro, hydroxymethyl, 1-hydroxyethyl,2-hydroxyethyl, 2-hydroxy-2-propyl, CF₃, OCF₃, SH, SCH₃, SOCH₃, SCF₃,COOH, COOCH₃, COCH₃, CH═O, acetoxy, amino, mono- or dialkylaminototalling 1-4 carbon atoms inclusive, acetamido, N-methylacetamido,carboxamido, N-alkylated carboxamido containing 1-4 carbon atomsinclusive, hydroxyacetyl and hydroxyacetoxy; and wherein for G¹ theoptional second and third rings may be fused to the first ring and/or toone another or may be separate rings connected to one another and/or tothe atom bearing G¹ by a single or double bond or by an interveningsubstituted or unsubstituted, linear or branched, saturated orunsaturated chain containing not more than 8 carbon atoms, not more than8 halogens, and not more than 4 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur; wherein E₁ is selected from═O, ═S, ═NH, ═NOR_(E) ⁸ where R_(E) ⁸ is hydrogen or a C₁ -C₈ normal orbranched alkyl radical, ═N--NH₂, hydrogen, halogen, --OH, --SH, --NH₂,--NH--NH₂, --N₃, --CN, --NO, --NO₂, --NHOH, --ONH₂, or is selected from:##STR56## wherein T¹ is selected from --O--, --S--, and --NH--; T² isselected from ═O, ═S, and ═N--R_(E) ⁶, in which R_(E) ⁶ may be hydrogen,hydroxy, cyano, or nitro; T^(2') is selected from ═O and ═S; T³, T⁴ andT^(4') are independently selected from --OH, --NH₂, --SH, --N₃,--NH--NH₂, and --NH--OR_(E) ⁷, in which R_(E) ⁷ may be hydrogen, C₁₋₃alkyl, or C₁₋₃ acyl; T³ may also be hydrogen or halogen; T⁵ -T^(5") areindependently selected from hydrogen and hydroxy; T⁵ may also behalogen; R_(E) ¹ is selected from hydrogen, halogen, hydroxy, nitro,nitroso, cyano, azide, --NH₂, --NH--OH, --SH, --O--NH₂, --NH--NH₂, --T¹--C(═T²)--T³, --C(═T²)--T³, --SiT⁵ T^(5') T^(5"), --T¹ --S(═O)(═T²)--T⁴,--S(═O)(═T²)--T⁴, --T¹ --P(--T⁴)--T^(4'), --P(--T⁴)--T^(4'), --T¹--P(═T^(2'))(--T³)--T⁴, and --P(═T^(2'))(--T³)--T⁴ ; R_(E) ² and R_(E) ³are individually selected from hydrogen, --C(═T²)--T³, cyano, nitro,azide, halogen and a C₁ -C₁₅ straight or branched chain, saturated,unsaturated and/or aromatic substituent optionally containing not morethan 10 halogen atoms and not more than 4 heteroatoms selected fromoxygen, nitrogen and sulfur; if R_(E) ¹ is cyano or C(═T²)--T³, thenR_(E) ² or R_(E) ³ may optionally be selected from --SiT⁵ T^(5') T^(5"),--T¹ --P(═T^(2'))(--T³)--T⁴, and --P(═T^(2'))(--T³)--T⁴ ; and R_(E) ⁴and R_(E) ⁵ are individually selected from hydrogen, halogen, cyano,nitro, --C(═T²)--T³, --T¹ --C(═T²)--T³, CR_(E) ¹ R_(E) ² R_(E) ³, --SiT⁵T^(5') T^(5"), --S(═O)(═T²)--T⁴, and --P(═T^(2'))(--T³)--T⁴ ; providedthat: the total P₁ ring skeleton may not comprise any of the followingsix minimum frameworks ##STR57## with or without additional ring(s)appended thereto.
 24. A composition of claim 23 wherein P₁ is P_(1R),##STR58## wherein carbons (1 and 2) or (2 and 3) may be joined by adouble bond; carbons (5 and 6) or (6 and 7) may be joined by a doublebond;S₁ E₁ may be bonded to carbon 5, 6 or 7; R_(A1) represents not morethan 8 identical or different substituents bonded independently viasingle and/or double bonds to carbons 1, 2 and/or 3, which substituentsmay independently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing groups, which halogens and groups taken togethercontain a total of not more than 30 carbon atoms, not more than 24halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur; R_(A3) represents not morethan 6 identical or different substituents bonded independently viasingle and/or double bonds to carbons 5, 6 and/or 7, which substituentsmay independently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated, and/or aromatic carbon-and/or heteroatom-containing groups, which halogens and groups takentogether contain not more than 12 carbon atoms, not more than 8 halogenatoms, and not more than 5 heteroatoms selected from oxygen, nitrogen,silicon, phosphorus and sulfur; and R_(A4) represents not more than 10identical or different substituents bonded independently via single ordouble bonds to carbons 9, 11, 12, 13 and/or 14, which substituents mayindependently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing groups, the group or groups optionally completing1-3 additional rings through bonds among themselves and/or 1-5additional rings when taken together with the 5-membered ring and itssubstituent(s) R_(A1), which halogen and groups taken together, includenot more than 50 carbon atoms, not more than 24 halogen atoms, and notmore than 15 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur.
 25. A composition of claim 24 wherein carbons 13and 14 of P_(1R) carry a substituted or unsubstituted cyclopropyl ring,forming P_(1P) ##STR59## wherein the R_(A5) and R_(A6) radicals mayindependently be hydrogen, halogen and/or a straight chain or branchedchain, cyclic or acyclic, saturated, unsaturated and/or aromatic carbon-and/or heteroatom-containing group, which halogens and groups takentogether contain a total of not more than 30 carbon atoms, not more than24 halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur.
 26. A composition of claim 25wherein the P_(1P) carbon 2 carries a methyl group, J¹ is hydroxy,carbon 9 carries a hydroxy group in the alpha configuration, carbons 10and 14 carry alpha hydrogens, carbon 11 carries a methyl group in thealpha configuration and R_(A5) and R_(A6) are both methyl, formingP_(1PP) ##STR60##
 27. A composition of claim 26 wherein S₁ E₁ takentogether comprise a saturated normal or branched C₂ -C₁₀ alkyl groupcontaining one hydroxy group separated from P₁ by not more than fourcarbon atoms.
 28. A composition of claim 23 wherein an orthoesterstructure is bonded to P_(1R) via one orthoester oxygen atom in thealpha configuration to carbon 9 and via two orthoester oxygen atoms inthe alpha configuration to any two of carbons 12-14, forming P_(1RR)##STR61## wherein R_(A7) represents hydrogen, halogen or a straightchain or branched chain, cyclic or acyclic, saturated, unsaturatedand/or aromatic carbon- and/or heteroatom-containing group, the groupoptionally completing 1 additional ring to carbon 1, which groupincludes not more than 30 carbon atoms, not more than 16 halogen atoms,and not more than 9 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur.
 29. A composition of claim 28 wherein the P_(1RR)carbon 2 carries a methyl group, J¹ is hydroxy, carbon 10 carries ahydrogen atom in the alpha configuration, carbon 11 carries a methylgroup in the alpha configuration and carbon 13 carries an isopropenylgroup, forming P_(1RRR) ##STR62##
 30. A composition of claim 23 whereinP₁ is P_(1I) ##STR63## wherein R_(A2I) represents not more than 8identical or different substituents bonded independently via singleand/or double bonds to carbons 1, 2 and/or 3, which substituents mayindependently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing groups, which halogens and groups taken togethercontain a total of not more than 30 carbon atoms, not more than 24halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur;R_(A3I) represents not morethan 6 identical or different substituents bonded independently viasingle and/or double bonds to carbons 5, 6 and/or 7, which substituentsmay independently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated, and/or aromatic carbon-and/or heteroatom-containing groups, which halogens and groups takentogether contain not more than 12 carbon atoms, not more than 8 halogenatoms, and not more than 5 heteroatoms selected from oxygen, nitrogen,silicon, phosphorus and sulfur; and R_(A4I) represents not more than 10identical or different substituents bonded independently via single ordouble bonds to carbons 9, 11, 12, 13 and/or 14, which substituents mayindependently be halogen(s) and/or straight chain or branched chain,cyclic or acyclic, saturated, unsaturated and/or aromatic carbon- and/orheteroatom-containing groups, the group or groups optionally completing1-3 additional rings through bonds among themselves and/or 1-5additional rings when taken together with the 5-membered ring and itssubstituent(s) R_(A2I), which halogen and groups taken together, includenot more than 50 carbon atoms, not more than 24 halogen atoms, and notmore than 15 heteroatoms selected from oxygen, nitrogen, silicon,phosphorus and sulfur.
 31. A composition of claim 30 wherein carbons 13and 14 of P_(1I) carry a substituted or unsubstituted cyclopropyl ring,forming P_(1IN) ##STR64## wherein the R_(A5I) and R_(A6I) radicals mayindependently be hydrogen, halogen and/or a straight chain or branchedchain, cyclic or acyclic, saturated, unsaturated and/or aromatic carbon-and/or heteroatom-containing group, which halogens and groups takentogether contain a total of not more than 30 carbon atoms, not more than24 halogen atoms and not more than 9 heteroatoms selected from oxygen,nitrogen, silicon, phosphorus and sulfur.
 32. A composition of claim 31wherein the P_(1IN) carbon 2 carries a methyl group, J¹ is hydroxy,carbon 11 carries a methyl group in the alpha configuration, carbon 14carries an alpha hydrogen and R_(A5I) and R_(A6I) are both methyl,forming P_(1INL) ##STR65##
 33. A protein kinase C-modulating compositionfor use in modulating the in vitro or in vivo activity of protein kinaseC, said composition comprising:a physiologically acceptablepharmaceutical carrier and a compound of claim 1 in a quantity ofbetween about 0.0001-1000 mg per unit dosage.
 34. A protein kinaseC-modulating composition for use in modulating the in vitro or in vivoactivity of protein kinase C, said composition comprising:an aqueous orpartly aqueous buffer or liquid carrier and a compound of claim 1 at aconcentration between about 0.1 pM to about 100 μM.
 35. A protein kinaseC-modulating composition for use in modulating the in vitro or in vivoactivity of protein kinase C, said composition comprising:aphysiologically acceptable pharmaceutical carrier and a compound ofclaim 2 in a quantity of between about 0.0001-1000 mg per unit dosage.36. A protein kinase C-modulating composition for use in modulating thein vitro or in vivo activity of protein kinase C, said compositioncomprising:an aqueous or partly aqueous buffer or liquid carrier and acompound of claim 2 at a concentration between about 0.1 pM to about 100μM.
 37. A protein kinase C-modulating composition for use in modulatingthe in vitro or in vivo activity of protein kinase C, said compositioncomprising:a physiologically acceptable pharmaceutical carrier and acompound of claim 3 in a quantity of between about 0.0001-1000 mg perunit dosage.
 38. A protein kinase C-modulating composition for use inmodulating the in vitro or in vivo activity of protein kinase C, saidcomposition comprising:an aqueous or partly aqueous buffer or liquidcarrier and a compound of claim 3 at a concentration between about 0.1pM to about 100 μM.
 39. A method of modulating protein kinase C activitycomprising the step of contacting protein kinase C, in vitro or in vivo,with a composition of claim
 33. 40. A method of modulating proteinkinase C activity comprising the step of contacting protein kinase C, invitro or in vivo, with a composition of claim
 36. 41. A method ofmodulating protein kinase C activity comprising the step of contactingprotein kinase C, in vitro or in vivo, with a composition of claim 35.42. A method of modulating protein kinase C activity comprising the stepof contacting protein kinase C, in vitro or in vivo, with a compositionof claim
 36. 43. A method of modulating protein kinase C activitycomprising the step of contacting protein kinase C, in vitro or in vivo,with a composition of claim
 57. 44. A method of modulating proteinkinase C activity comprising the step of contacting protein kinase C, invitro or in vivo, with a composition of claim
 38. 45. A method oftreating inflammation in a mammal in need of anti-inflammatory treatmentcomprising the step of contacting said mammal with a composition ofclaim
 21. 46. A method of treating inflammation in a mammal in need ofanti-inflammatory treatment comprising the step of contacting saidmammal with a composition of claim
 22. 47. A method of treatinginflammation in a mammal in need of anti-inflammatory treatmentcomprising the step of contacting said mammal with a composition ofclaim
 23. 48. A method of treating inflammation in a mammal in need ofanti-inflammatory treatment comprising the step of contacting saidmammal with a composition of claim
 24. 49. A method of treatinginflammation in a mammal in need of anti-inflammatory treatmentcomprising the step of contacting said mammal with a composition ofclaim
 25. 50. A method of treating inflammation in a mammal in need ofanti-inflammatory treatment comprising the step of contacting saidmammal with a composition of claim
 26. 51. A method of treatinginflammation in a mammal in need of anti-inflammatory treatmentcomprising the step of contacting said mammal with a composition ofclaim
 27. 52. A method of treating inflammation in a mammal in need ofanti-inflammatory treatment comprising the step of contacting saidmammal with a composition of claim
 28. 53. A method of treatinginflammation in a mammal in need of anti-inflammatory treatmentcomprising the step of contacting said mammal with a composition ofclaim
 29. 54. A method of treating inflammation in a mammal in need ofanti-inflammatory treatment comprising the step of contacting saidmammal with a composition of claim
 30. 55. A method of treatinginflammation in a mammal in need of anti-inflammatory treatmentcomprising the step of contacting said mammal with a composition ofclaim
 31. 56. A method of treating inflammation in a mammal in need ofanti-inflammatory treatment comprising the step of contacting saidmammal with a composition of claim 32.